Sample records for larval mosquitoes aedes

Full Text Available The mosquito-borne La Crosse virus (LACV; Family Bunyaviridae may cause encephalitis, primarily in children, and is distributed throughout much of the eastern United States. No antivirals or vaccines are available for LACV, or most other mosquito-borne viruses, and prevention generally relies on mosquito control. We sought to determine whether coffee extracts could interfere with LACV replication and vector mosquito development. Both regular and decaffeinated coffee demonstrated significant reductions in LACV replication in direct antiviral assays. This activity was not due to the presence of caffeine, which did not inhibit the virus life cycle. Aedes albopictus (Skuse (Diptera: Culicidae mosquito larvae suffered near total mortality when reared in high concentrations of regular and decaffeinated coffee and in caffeine. Following larval exposure to sublethal coffee concentrations, adult Ae. albopictus mosquitoes had signficantly reduced whole-body LACV titers five days post-infection, compared to larvae reared in distilled water. These results suggest that it may be possible to both control mosquito populations and alter the vector competence of mosquitoes for arthropod-borne viruses by introducing antiviral compounds into the larval habitat.

With field releases starting in Brazil, particular interest must be given to understanding how the endosymbiotic bacterium Wolbachia pipientis affects Aedes aegypti mosquitoes with a Brazilian genetic background. Currently, there is limited information on how the bacterium affects phenotypic traits such as larval development rate, metabolic reserves and morphometric parameters in Ae. aegypti. Here, we analyze for the first time, the effect of Wolbachia on these key phenotypes and consider how this might impact the potential of the bacterium as a disease control agent in Brazil. We examined the influence of the wMel strain of Wolbachia in laboratory Ae. aegypti with a Brazilian genetic background, reared under different larval densities. Pupae formation was counted daily to assess differences in development rates. Levels of metabolic reserves and morphometric parameters were assessed in adults resulting from each larval condition. wMel infection led to more rapid larval development at higher densities for both males and females, with no effect under less crowded conditions in females. Infection also led to reduced body size at both high and low density, but not at intermediate density, although the scale of this difference was maintained regardless of larval density, in comparison to uninfected individuals. Wing shape also varied significantly between infected and uninfected mosquitoes due to larval density. Glycogen levels in uninfected mosquitoes decreased under higher larval density, but were consistently high with Wolbachia infection, regardless of larval density. We demonstrate that the wMel Wolbachia strain can positively influence some important host fitness traits, and that this interaction is directly linked to the conditions in which the host is reared. Combined with previously published data, these results suggest that this Wolbachia strain could be successfully used as part of the Eliminate Dengue Program in Brazil.

Density-dependent processes such as larval competition may be important regulatory factors among some mosquito species. The application of pesticides used for control may alter these density-dependent interactions with consequences for the number of survivors and associated sublethal and chronic effects on these individuals. We examined how intraspecific competition among larvae and low concentrations of malathion alter Aedes aegypti L. and Aedes albopictus Skuse adult life history traits and competence for arboviruses using Sindbis virus as a model system. Larvae were reared at densities of 150 and 300 larvae per container and in the absence or presence of 0.04 parts per million of malathion, before surviving females were exposed to an infectious blood meal containing 10(5) plaque-forming units/ml Sindbis virus. For both species, competition and the presence of malathion reduced survival to adulthood. The presence of malathion eliminated the negative effects of competition that resulted in lengthened development time and smaller-sized adults. For Ae. aegypti, but not Ae. albopictus, high competition conditions and the presence of malathion independently and not interactively led to an increase in virus dissemination from the midgut. Our results suggest that larval competition and chemical contaminants may influence disease transmission directly by altering adult mosquito fitness and indirectly by altering vector interactions with arboviruses.

As the relationship between a given life-history trait and fitness is not necessarily the same for the two sexes, an 'intersexual ontogenetic conflict' may arise. We analysed the phenotypic reaction to intraspecific larval competition of the mosquito, Aedes aegypti, asking: (i) Do both sexes pay the cost of competition with the same life-history traits and are they equal competitors? (ii) Is there a specific cost of competition beyond sharing food resources? We found that competition incurs a specific cost that was expressed differently by the two sexes. Indeed, each sex maintained the more important life-history trait(s) for their fitness (developmental time for males and body weight and size for females) at the expense of other traits, thus minimizing the effects of competition on their fitness. The competition exerted by females was estimated as being more intense, probably linked with the greater importance of body size for their fitness.

The larvae of the mosquito, Aedes aegypti normally inhabit freshwater (FW) where they face dilution of body fluids by osmotic influx of water. In response, the physiological actions of the anal papillae result in ion uptake while the Malpighian tubules and rectum work in concert to excrete excess water. In an apparent paradox, the anal papillae express aquaporins (AQPs) and are sites of water permeability which, if AQPs are expressed by the epithelium, apparently exaggerates the influx of water from their dilute environment. Recently, naturally breeding populations of A. aegypti were found in brackish water (BW), an environment which limits the osmotic gradient. Given that salinization of FW is an emerging environmental issue and that these larvae would presumably need to adjust to these changing conditions, this study investigates the expression of AQPs in the anal papillae and their response to rearing in hypo-osmotic and near isosmotic conditions. Transcripts of all six Aedes AQP homologs were detectable in the anal papillae and the transcript abundance of three AQP homologs in the papillae was different between rearing conditions. Using custom made antibodies, expression of two of these AQP homologs (AQP4 and AQP5) was localized to the syncytial epithelium of the anal papillae. Furthermore, the changes in transcript abundance of these two AQPs between the rearing conditions, were manifested at the protein level. Results suggest that AQP4 and AQP5 play an important physiological role in larval responses to changes in environmental salinity.

As the fat body is a critical tissue for mosquito development, metamorphosis, immune and reproductive system function, characterization of regulatory modules targeting gene expression to the female mosquito fat body at distinct life stages is much needed for multiple, varied strategies for controlling vector-borne diseases such as dengue and malaria. The hexameric storage protein, Hexamerin-1.2, of the mosquito, Aedes atropalpus, is female-specific and uniquely expressed in the fat body of fo...

container-in- habiting mosquitoes that transmit disease agents, outcompete native species, and continue to expand their range in theUnited States. Both...days to complete evaporation in containers in- habited by the most common species collected, us- ing themean temperature of 18.6Cduring sampling... snack bags, and Sty- rofoam cups and bowls. Evaporation analyses showed that Culex species were more common in containers that are less inßuenced by

As the fat body is a critical tissue for mosquito development, metamorphosis, immune and reproductive system function, the characterization of regulatory modules targeting gene expression to the female mosquito fat body at distinct life stages is much needed for multiple, varied strategies for controlling vector-borne diseases such as dengue and malaria. The hexameric storage protein, Hexamerin-1.2, of the mosquitoAedes atropalpus is female-specific and uniquely expressed in the fat body of fourth instar larvae and young adults. We have identified in the Hex-1.2 gene, a short regulatory module that directs female-, tissue-, and stage-specific lacZ reporter gene expression using a heterologous promoter in transgenic lines of the dengue vector Aedes aegypti. Male transgenic larvae and pupae of one line expressed no Escherichia coli β-galactosidase or transgene product; in two other lines reporter gene activity was highly female-biased. All transgenic lines expressed the reporter only in the fat body; however, lacZ mRNA levels were no different in males and females at any stage examined, suggesting that the gene regulatory module drives female-specific expression by post-transcriptional regulation in the heterologous mosquito. This regulatory element from the Hex-1.2 gene thus provides a new molecular tool for transgenic mosquito control as well as functional genetic analysis in aedine mosquitoes.

Adult Aedes aegypti (Linnaeus) (Diptera: Culicidae) were previously recovered from emergence traps on septic tanks in southeastern Puerto Rico. In this study we quantified immature mosquito abundance and its relationship with structural variables of the septic tanks and chemical properties of the water containing raw sewage. A miniaturized floating funnel trap was used to sample 89 septic tanks for larvae in the Puerto Rican community of Playa-Playita. Aedes aegypti larvae were recovered from 18% of the sampled tanks (10.3 larvae per septic tank per day). Larval presence was positively associated with cracking of the septic tank walls and uncovered access ports. Larval abundance was positively associated with cracking of the septic tank walls and larger tank surface areas, and inversely associated with the total dissolved solids (TDS). Culex quinquefasciatus (Say) larvae were also recovered from 74% of the septic tanks (129.6 larvae per septic tank per day). Larval presence was negatively associated with TDS in the water and larval abundance was positively associated with cracking of the septic tank walls. A screened, plastic emergence trap was used to sample 93 septic tanks within the community for Ae. aegypti and Cx. quinquefasciatus adults. Aedes aegypti adults were recovered from 49% of the sampled tanks (8.7 adults per septic tank per day) and Cx. quinquefasciatus adults were recovered from 97% of the sampled tanks (155.5 adults per septic tank per day). Aedes aegypti adult presence was positively associated with cracking, uncapped openings and septic water pH. The Ae. aegypti adult counts were positively associated with cracking and inversely associated with TDS and conductivity. This study marks the first published record of the recovery of Ae. aegypti larvae from holding tanks containing raw sewage in the Caribbean region. Our study indicates that Ae. aegypti larvae are present in sewage water and that septic tanks have at least the potential to maintain

Full Text Available Abstract Background Oviposition-site choice is an essential component of the life history of all mosquito species. According to the oviposition-preference offspring-performance (P-P hypothesis, if optimizing offspring performance and fitness ensures high overall reproductive fitness for a given species, the female should accurately assess details of the heterogeneous environment and lay her eggs preferentially in sites with conditions more suitable to offspring. Methods We empirically tested the P-P hypothesis using the mosquito species Aedes albopictus by artificially manipulating two habitat conditions: diet (measured as mg of food added to a container and conspecific density (CD; number of pre-existing larvae of the same species. Immature development (larval mortality, development time to pupation and time to emergence and fitness (measured as wing length were monitored from first instar through adult emergence using a factorial experimental design over two ascending gradients of diet (2.0, 3.6, 7.2 and 20 mg food/300 ml water and CD (0, 20, 40 and 80 larvae/300 ml water. Treatments that exerted the most contrasting values of larval performance were recreated in a second experiment consisting of single-female oviposition site selection assay. Results Development time decreased as food concentration increased, except from 7.2 mg to 20.0 mg (Two-Way CR ANOVA Post-Hoc test, P > 0.1. Development time decreased also as conspecific density increased from zero to 80 larvae (Two-Way CR ANOVA Post-Hoc test, P . Combined, these results support the role of density-dependent competition for resources as a limiting factor for mosquitolarval performance. Oviposition assays indicated that female mosquitoes select for larval habitats with conspecifics and that larval density was more important than diet in driving selection for oviposition sites. Conclusions This study supports predictions of the P-P hypothesis and provides a mechanistic understanding

The management and control of mosquito vectors of human disease currently rely primarily on chemical insecticides. However, larvicidal treatments can be effective, and if based on biological insecticides, they can also ameliorate the risk posed to human health by chemical insecticides. The aerobic bacteria Bacillus thuringiensis and Lysinibacillus sphaericus have been used for vector control for a number of decades. But a more cost-effective use would be an anaerobic bacterium because of the ease with which these can be cultured. More recently, the anaerobic bacterium Clostridium bifermentans subsp. malaysia has been reported to have high mosquitocidal activity, and a number of proteins were identified as potentially mosquitocidal. However, the cloned proteins showed no mosquitocidal activity. We show here that four toxins encoded by the Cry operon, Cry16A, Cry17A, Cbm17.1, and Cbm17.2, are all required for toxicity, and these toxins collectively show remarkable selectivity for Aedes rather than Anopheles mosquitoes, even though C. bifermentans subsp. malaysia is more toxic to Anopheles. Hence, toxins that target Anopheles are different from those expressed by the Cry operon. PMID:25002432

An integrated larvalmosquito control program was carried out in Tiputa village on Rangiroa atoll of French Polynesia. Mosquito abundance before and after treatment was compared with the abundance in an untreated village. Mosquitolarval habitats consisted of large concrete or polyurethane cisterns, wells, and 200-liter drums. Depending on the target species, larval habitat category, its configuration, and purpose (drinking consumption or not), abatement methods consisted of sealing the larval habitats with mosquito gauze, treating them with 1% Temephos, covering the water with a 10-cm thick layer of polystyrene beads or introducing fish (Poecillia reticulata Rosen & Bailey). All premises of the chosen village were treated and a health education program explained basic mosquito ecology and the methods of control. A community health agent was trained to continue the control program at the end of the experiment. Entomological indices from human bait collections and larval surveys indicated that mosquito populations were reduced significantly, compared with concurrent samples from the untreated control village, and that mosquito control remained effective for 6 mo after treatment. Effects of the treatment were noticed by the inhabitants in terms of a reduction in the number of mosquito bites. In the Polynesian context, such control programs may succeed in the long-term only if strong political decisions are taken at the village level, if a community member is designated as being responsible for maintaining the program, and if the inhabitants are motivated sufficiently by the mosquito nuisance to intervene.

Full Text Available Failure in detecting naturally occurring breeding sites of Aedesmosquitoes can bias the conclusions drawn from field studies, and hence, negatively affect intervention outcomes. We characterized the habitats of immature Aedesmosquitoes and explored species dynamics along a rural-to-urban gradient in a West Africa setting where yellow fever and dengue co-exist.Between January 2013 and October 2014, we collected immature Aedesmosquitoes in water containers in rural, suburban, and urban areas of south-eastern Côte d'Ivoire, using standardized sampling procedures. Immature mosquitoes were reared in the laboratory and adult specimens identified at species level.We collected 6,159, 14,347, and 22,974 Aedesmosquitoes belonging to 17, 8, and 3 different species in rural, suburban, and urban environments, respectively. Ae. aegypti was the predominant species throughout, with a particularly high abundance in urban areas (99.374%. Eleven Aedeslarval species not previously sampled in similar settings of Côte d'Ivoire were identified: Ae. albopictus, Ae. angustus, Ae. apicoargenteus, Ae. argenteopunctatus, Ae. haworthi, Ae. lilii, Ae. longipalpis, Ae. opok, Ae. palpalis, Ae. stokesi, and Ae. unilineatus. Aedes breeding site positivity was associated with study area, container type, shade, detritus, water turbidity, geographic location, season, and the presence of predators. We found proportionally more positive breeding sites in urban (2,136/3,374, 63.3%, compared to suburban (1,428/3,069, 46.5% and rural areas (738/2,423, 30.5%. In the urban setting, the predominant breeding sites were industrial containers (e.g., tires and discarded containers. In suburban areas, containers made of traditional materials (e.g., clay pots were most frequently encountered. In rural areas, natural containers (e.g., tree holes and bamboos were common and represented 22.1% (163/738 of all Aedes-positive containers, hosting 18.7% of the Aedes fauna. The predatory mosquito

Failure in detecting naturally occurring breeding sites of Aedesmosquitoes can bias the conclusions drawn from field studies, and hence, negatively affect intervention outcomes. We characterized the habitats of immature Aedesmosquitoes and explored species dynamics along a rural-to-urban gradient in a West Africa setting where yellow fever and dengue co-exist. Between January 2013 and October 2014, we collected immature Aedesmosquitoes in water containers in rural, suburban, and urban areas of south-eastern Côte d'Ivoire, using standardized sampling procedures. Immature mosquitoes were reared in the laboratory and adult specimens identified at species level. We collected 6,159, 14,347, and 22,974 Aedesmosquitoes belonging to 17, 8, and 3 different species in rural, suburban, and urban environments, respectively. Ae. aegypti was the predominant species throughout, with a particularly high abundance in urban areas (99.374%). Eleven Aedeslarval species not previously sampled in similar settings of Côte d'Ivoire were identified: Ae. albopictus, Ae. angustus, Ae. apicoargenteus, Ae. argenteopunctatus, Ae. haworthi, Ae. lilii, Ae. longipalpis, Ae. opok, Ae. palpalis, Ae. stokesi, and Ae. unilineatus. Aedes breeding site positivity was associated with study area, container type, shade, detritus, water turbidity, geographic location, season, and the presence of predators. We found proportionally more positive breeding sites in urban (2,136/3,374, 63.3%), compared to suburban (1,428/3,069, 46.5%) and rural areas (738/2,423, 30.5%). In the urban setting, the predominant breeding sites were industrial containers (e.g., tires and discarded containers). In suburban areas, containers made of traditional materials (e.g., clay pots) were most frequently encountered. In rural areas, natural containers (e.g., tree holes and bamboos) were common and represented 22.1% (163/738) of all Aedes-positive containers, hosting 18.7% of the Aedes fauna. The predatory mosquito species

The Asian tiger mosquito, Aedes albopictus (Skuse), is a highly invasive container-inhabiting species with a global distribution. This mosquito, similar to other Stegomyia species such as Aedes aegypti (L.), is highly adapted to urban and suburban areas, and commonly oviposits in artificial containers, which are ubiquitous in these peridomestic environments. The increase in speed and amount of international travel and commerce, coupled with global climate change, have aided in the resurgence and expansion of Stegomyia species into new areas of North America. In many parts of their range, both species are implicated as significant vectors of emerging and re-emerging arboviruses such as dengue, chikungunya, and now Zika. Although rapid and major advances have been made in the field of biology, ecology, genetics, taxonomy, and virology, relatively little has changed in the field of mosquito control in recent decades. This is particularly discouraging in regards to container-inhabiting mosquitoes, because traditional integrated mosquito management (IMM) approaches have not been effective against these species. Many mosquito control programs simply do not possess the man-power or necessary financial resources needed to suppress Ae. albopictus effectively. Therefore, control of mosquito larvae, which is the foundation of IMM approaches, is exceptionally difficult over large areas. This review paper addresses larval habitats, use of geographic information systems for habitat preference detection, door-to-door control efforts, source reduction, direct application of larvicides, biological control agents, area-wide low-volume application of larvicides, hot spot treatments, autodissemination stations, public education, adult traps, attractive-toxic sugar bait methods, lethal ovitraps, barrier-residual adulticides, hand-held ultra-low-volume adulticides, area-wide adulticides applied by ground or air, and genetic control methods. The review concludes with future

Introduction: The life history of Aedes aegypti presents diverse challenges to its diuretic system. During the larval and pupal life stages mosquitoes are aquatic. With the emergence of the adult they become terrestrial. This shifts the organism within minutes from an aquatic environment to a terrestrial environment where dehydration has to be avoided. In addition, female mosquitoes take large blood meals, which present an entirely new set of challenges to salt and water homeostasis. Methods: To determine differences in gene expression associated with these different life stages, we performed an RNA-seq analysis of the main diuretic tissue in A. aegypti, the Malpighian tubules. We compared transcript abundance in 4th instar larvae to that of adult females and analyzed the data with a focus on transcripts that encode proteins potentially involved in diuresis, like water and solute channels as well as ion transporters. We compared our results against the model of potassium- and sodium chloride excretion in the Malpighian tubules proposed by Hine et al. (2014), which involves at least eight ion transporters and a proton-pump. Results: We found 3,421 of a total number of 17,478 (19.6%) unique transcripts with a P aegypti mosquitoes, highlighting key differences in their renal systems that arise as they transform from an aquatic filter-feeding larval stage to a terrestrial, blood-feeding adult stage.

Dengue is the most important arboviral disease worldwide and the principal vector-borne disease in Costa Rica. Control of Aedes aegypti populations through source reduction is still considered the most effective way of prevention and control, although it has proven ineffective or unsustainable in many areas with a history of mosquito control. In this study, seasonal profiles and productivity of Aedes aegypti were analyzed in the city of Puntarenas, Costa Rica, where vector control has been practiced for more than ten years. Households contained more than 80% of larval habitats identified, although presence of habitats was more likely in other locations like lots and streets. In the wet season, habitats in the "other" category, like appliances, small manholes, and miscellaneous containers, were the most frequent habitats observed as well as the most common and productive habitats for Ae. aegypti. In the dry season, domestic animal drinking containers were very common, although concrete washtubs contained 79% of Ae. aegypti pupae collected. Individually, non-disposable habitats were as likely or more likely to contain mosquito larvae, and large containers were more likely to harbor mosquito larvae than the small ones only in the dry season. Considering various variables in the logistic regressions, predictors for Ae. aegypti in a habitat were habitat type (p aegypti larval indices and pupal indices in Puntarenas were high enough to allow viral transmission during the wet season. In spite of continued vector control, it has not been possible to reduce vector densities below threshold levels in Puntarenas, and the habitat profiles show that non-household locations, as well as non-disposable containers, should be targeted in addition to the standard control activities.

Full Text Available Introduction: The life history of Aedes aegypti presents diverse challenges to its diuretic system. During the larval and pupal life stages mosquitoes are aquatic. With the emergence of the adult they become terrestrial. This shifts the organism within minutes from an aquatic environment to a terrestrial environment where dehydration has to be avoided. In addition, female mosquitoes take large blood meals, which present an entirely new set of challenges to salt and water homeostasis.Methods: To determine differences in gene expression associated with these different life stages, we performed an RNA-seq analysis of the main diuretic tissue in A. aegypti, the Malpighian tubules. We compared transcript abundance in 4th instar larvae to that of adult females and analyzed the data with a focus on transcripts that encode proteins potentially involved in diuresis, like water and solute channels as well as ion transporters. We compared our results against the model of potassium- and sodium chloride excretion in the Malpighian tubules proposed by Hine et al. (2014, which involves at least eight ion transporters and a proton-pump.Results: We found 3,421 of a total number of 17,478 (19.6% unique transcripts with a P < 0.05 and at least a 2.5 fold change in expression levels between the two groups. We identified two novel transporter genes that are highly expressed in the adult Malpighian tubules, which have not previously been part of the transport model in this species and may play important roles in diuresis. We also identified candidates for hypothesized sodium and chloride channels. Detoxification genes were generally higher expressed in larvae.Significance: This study represents the first comparison of Malpighian tubule transcriptomes between larval and adult A. aegypti mosquitoes, highlighting key differences in their renal systems that arise as they transform from an aquatic filter-feeding larval stage to a terrestrial, blood-feeding adult stage.

97% of the total are Aedes, Anopheles, and Culex , and a good series of all stages (male, female, larva, and pupa ) were obtained for most species of...Is. SUPPLEMENTARY NOTES 19. KEY WORDS (Continue on reverse side If necessary and tdentify by block number) Mosquitoes Africa Aedes Culicidae...entomologists with keys , necessary literature, and other identification guides, to the loan of specialized .0 collecting and rearing equipment, which cannot be

Because of increasing insecticide resistance, new pesticides are needed. Flowering plants have been the source of useful pesticides in the past. We studied 15 chemicals isolated from a poisonous pasture plant for activity against the yellow fever mosquito. We found that dehydrotremetone was effectiv...

To determine the potential role of vertical transmission in Zika virus expansion, we evaluated larval pools of perorally infected Aedes aegypti and Ae. albopictus adult female mosquitoes; ≈1/84 larvae tested were Zika virus-positive; and rates varied among mosquito populations. Thus, vertical transmission may play a role in Zika virus spread and maintenance.

Full Text Available Aquatic environments can be restricted with the amount of available food resources especially with changes to both abiotic and biotic conditions. Mosquito larvae, in particular, are sensitive to changes in food resources. Resource limitation through inter-, and intra-specific competition among mosquitoes are known to affect both their development and survival. However, much less is understood about the effects of non-culicid controphic competitors (species that share the same trophic level. To address this knowledge gap, we investigated and compared mosquitolarval development, survival and adult size in two experiments, one with different densities of non-culicid controphic conditions and the other with altered resource conditions. We used Aedes camptorhynchus, a salt marsh breeding mosquito and a prominent vector for Ross River virus in Australia. Aedes camptorhynchus usually has few competitors due to its halo-tolerance and distribution in salt marshes. However, sympatric ostracod micro-crustaceans often co-occur within these salt marshes and can be found in dense populations, with field evidence suggesting exploitative competition for resources. Our experiments demonstrate resource limiting conditions caused significant increases in mosquito developmental times, decreased adult survival and decreased adult size. Overall, non-culicid exploitation experiments showed little effect on larval development and survival, but similar effects on adult size. We suggest that the alterations of adult traits owing to non-culicid controphic competition has potential to extend to vector-borne disease transmission.

Objective:To determine the larval habitats of mosquito fauna and possible impact of land use/land cover changes on the epidemiology of mosquito-borne diseases in Osogbo metropolis, Southwestern, Nigeria. Methods: All accessible larval habitats were surveyed between May and September, 2011 in Osogbo metropolis while Land Use/ Land cover of the city was analyzed using 2 Lansat Multispectral Scanner satellite imagery of SPOT 1986 and LANDSAT TM 2009. Results:A total of six species namely, Aedes aegypti, Aedes albopictus, Aedes vittatus, Anopheles gambiae complex, Culex quinquefasciatus and Eretmapodite chrysogaster were encountered during the study. The occurrence and contribution of disused tyres was significantly higher (P0.05). The accessible land use/land covered of the study area between 1986 and 2009 showed that the wet land coverage and settlement area increased from 0.19 to 9.09 hectare and 1.00 to 2.01 hectare respectively while the forest area decreased from 60.18 to 50.14 hectare. Conclusion: The contribution of the habitats coupled with the increasing rate of flooded environment which could provide ample breeding sites for mosquitoes call for sustained environmental sanitation and management in Osogbo metropolis.

Full Text Available Expression of sterol carrier protein-2 (SCP-2 in Aedes aegypti shows a distinct temporal/spatial pattern throughout the life cycle. In order to identify the transcription factors responsible for the larval temporal/spatial regulation of AeSCP-2 transcription, AeSCP-2 promoter activities were studied in vivo via transient transfection of promoter/reporter gene assays. Regulatory sequences upstream -1.3 kb of the transcription start site of AeSCP-2 were found to be critical for the in vivo temporal/spatial promoter activity. Interestingly, the -1.6 kb promoter sequence efficiently drove the larval midgut-specific siRNA expression, indicating that the -1.6 kb upstream sequence is sufficient for temporal/spatial AeSCP-2 transcriptional activity. Four transcription factors were identified in the midgut nuclear extract from feeding larvae via labeled -1.6/-1.3 kb DNA probe pull-down and proteomic analysis. Co-transfection of the promoter/reporter gene with inducible siRNA expression of each transcription factor was performed to confirm the regulatory function of individual transcription factor on AeSCP-2 transcriptional activities in the larval midgut. The results indicate that two of the identified transcription factors, Thanatos-associated protein (THAP and activating transcription factor-2 (ATF-2, antagonistically control AeSCP-2 transcriptional activity in the midgut of feeding larvae via the regulatory sequences between -1.6 to -1.3 kb 5' upstream of the transcription start site. In vivo expression knockdown of THAP and ATF-2 resulted in significant changes in developmental progression, which may be partially due to their effects on AeSCP-2 expression.

Expression of sterol carrier protein-2 (SCP-2) in Aedes aegypti shows a distinct temporal/spatial pattern throughout the life cycle. In order to identify the transcription factors responsible for the larval temporal/spatial regulation of AeSCP-2 transcription, AeSCP-2 promoter activities were studied in vivo via transient transfection of promoter/reporter gene assays. Regulatory sequences upstream −1.3 kb of the transcription start site of AeSCP-2 were found to be critical for the in vivo temporal/spatial promoter activity. Interestingly, the −1.6 kb promoter sequence efficiently drove the larval midgut-specific siRNA expression, indicating that the −1.6 kb upstream sequence is sufficient for temporal/spatial AeSCP-2 transcriptional activity. Four transcription factors were identified in the midgut nuclear extract from feeding larvae via labeled −1.6/−1.3 kb DNA probe pull-down and proteomic analysis. Co-transfection of the promoter/reporter gene with inducible siRNA expression of each transcription factor was performed to confirm the regulatory function of individual transcription factor on AeSCP-2 transcriptional activities in the larval midgut. The results indicate that two of the identified transcription factors, Thanatos-associated protein (THAP) and activating transcription factor-2 (ATF-2), antagonistically control AeSCP-2 transcriptional activity in the midgut of feeding larvae via the regulatory sequences between −1.6 to −1.3 kb 5′ upstream of the transcription start site. In vivo expression knockdown of THAP and ATF-2 resulted in significant changes in developmental progression, which may be partially due to their effects on AeSCP-2 expression. PMID:23056538

As a geographically isolated island chain with no native mosquitoes, Hawaii is a model for examining the mechanisms behind insect vector invasions and their subsequent interactions with each other and with human populations. The yellow fever mosquito, Aedes aegypti, and the Asian tiger mosquito, Ae. albopictus, have been responsible for epidemics of dengue in Hawaii. As one of the world's earliest locations to be invaded by both species, Hawaii's history is particularly relevant because both species are currently invading new areas worldwide and are implicated in outbreaks of emergent or reemergent pathogens such as dengue, chikungunya, and yellow fever. Here we analyze the historical records of mosquito introductions in order to understand the factors that have led to the current distribution of these 2 mosquitoes in the Hawaiian Islands.

High speed video observations of free flying male Aedes aegypti mosquitoes, the dengue and yellow fever vector, along with custom measurement methods, enable measurement of wingbeat frequency, body position and body orientation of mosquitoes during flight. We find these mosquitoes flap their wings at approximately 850 Hz. We also generate body yaw, body pitch and wing deviation measurements with standard deviations of less than 1 degree and find that sideways velocity and acceleration are important components of mosquito motion. Rapid turns involving changes in flight direction often involve large sideways accelerations. These do not correspond to commensurate changes in body heading, and the insect's flight direction and body heading are decoupled during flight. These findings call in to question the role of yaw control in mosquito flight. In addition, using orientation data, we find that sideways accelerations are well explained by roll-based rotation of the lift vector. In contrast, the insect's body pitch...

Aedes aegypti, the principal vector of dengue fever, have been genetically engineered for use in a sterile insect control programme. To improve our understanding of the dispersal ecology of mosquitoes and to inform appropriate release strategies of 'genetically sterile' male Aedes aegypti detailed knowledge of the dispersal ability of the released insects is needed. The dispersal ability of released 'genetically sterile' male Aedes aegypti at a field site in Brazil has been estimated. Dispersal kernels embedded within a generalized linear model framework were used to analyse data collected from three large scale mark release recapture studies. The methodology has been applied to previously published dispersal data to compare the dispersal ability of 'genetically sterile' male Aedes aegypti in contrasting environments. We parameterised dispersal kernels and estimated the mean distance travelled for insects in Brazil: 52.8 m (95% CI: 49.9 m, 56.8 m) and Malaysia: 58.0 m (95% CI: 51.1 m, 71.0 m). Our results provide specific, detailed estimates of the dispersal characteristics of released 'genetically sterile' male Aedes aegypti in the field. The comparative analysis indicates that despite differing environments and recapture rates, key features of the insects' dispersal kernels are conserved across the two studies. The results can be used to inform both risk assessments and release programmes using 'genetically sterile' male Aedes aegypti.

Background: We aimed to extract the ingredients from leaves of Gossypium hirsutum (Bt cotton) using different solvents and evaluate for potential use to control different larval stages of mosquito species, Aedes aegypti and Anopheles stephensi. Methods: Qualitative and quantitative estimation of ingredients from Go. hirsutum (Bt) plant extract was carried out and their inhibitory action against mosquito larvae was determined using mosquito larvicidal assay. Results: LC50 values of water, etha...

Full Text Available Wild female Aedes aegypti and Ae. albopictus were allowed to lay eggs in (i ovitraps with different concentrations of NaCl, (ii different coloured ovistrips, (iii water from different sources, (iv larva holding water, and different sized ovitraps for oviposition preference. Oviposition cycle was also studied in different photoperiod regimens. The number of eggs laid was observed to gradually decrease with increase in NaCl concentration in both the species. Experiments were conducted to determine egg laying preference for any specific colour of the ovistrip and black ovistrip was found to be most preferred by both the species. For oviposition preference, eight water samples collected from different sources were used and it was observed that the maximum number of eggs was laid in ovitraps containing distilled water followed by tap water. In addition, Aedesmosquitoes laid more number of eggs in ovitraps containing larval holding water than ovitraps containing distilled water. Further, both the species did not lay any egg in the smallest used ovitrap although the number of eggs was maximally deposited in the largest ovitrap used. In the present studies, both the Aedes species laid the maximum number of eggs in the 4th quarter of the light period with normal 12 h light and dark phases (LD 12 : 12.

Darwin's northern suburbs border an extensive coastal reed and upper mangrove wetland recognized as an important larval habitat for Aedes vigilax (Skuse), the northern salt marsh mosquito, an established vector for Ross River and Barmah Forest viruses and an appreciable pest species. We sought to identify the most important vegetation categories associated with Ae. vigilax breeding to maximize the efficiency of mosquito control efforts. Using a generalized linear model with negative binominal distribution and log link, this study compares larval densities, determined by focused dipping, between 13 discernable vegetation categories. The incidence rate ratios (RR) generated can be used to compare the magnitude of larval densities for each vegetation category, compared with the reference category. Aedes vigilax larval densities were almost ten times greater in artificial drainage areas (RR=9.82), followed by tide-affected reticulate (Sporobolus/Xerochloa) areas (RR=8.15), then Schoenoplectus/mangroves (RR=2.29), compared with the reference vegetation category "lower mangroves." Furthermore, larval densities were highest in May, due to tidal inundation, for drainage areas and tide-affected reticulates (RR=12.2, 11.7, respectively) compared with March, the reference month. Thus, to maximize the efficiency of aerial salt marsh mosquito control operations in this wetland, larval control is best accomplished by concentrating on drains, Schoenoplectus/mangroves, and tide-affected reticulate areas, commencing early after the wet season. These results should apply to other areas of salt marsh mosquito breeding across northern Australia.

Despite the growing awareness that larval competition can influence adult mosquito life history traits including susceptibility to pathogens, the net effect of larval competition on human risk of exposure to mosquito-borne pathogens remains poorly understood. We examined how intraspecific larval competition affects dengue-2 virus (DENV-2) extrinsic incubation period and vectorial capacity of its natural vector Aedes albopictus. Adult Ae. albopictus from low and high-larval density conditions were orally challenged with DENV-2 and then assayed for virus infection and dissemination rates following a 6, 9, or 12-day incubation period using real-time quantitative reverse transcription PCR. We then modeled the effect of larval competition on vectorial capacity using parameter estimates obtained from peer-reviewed field and laboratory studies. Larval competition resulted in significantly longer development times, lower emergence rates, and smaller adults, but did not significantly affect the extrinsic incubation period of DENV-2 in Ae. albopictus. Our vectorial capacity models suggest that the effect of larval competition on adult mosquito longevity likely has a greater influence on vectorial capacity relative to any competition-induced changes in vector competence. Furthermore, we found that large increases in the viral dissemination rate may be necessary to compensate for small competition-induced reductions in daily survivorship. Our results indicate that mosquito populations that experience stress from larval competition are likely to have a reduced vectorial capacity, even when susceptibility to pathogens is enhanced.

Full Text Available Background & objectives: The district of Thiruvananthapuram reports the maximum number of cases of dengue in the state of Kerala. To determine the larval diversity, density and breeding site preferences of Aedesmosquitoes, during pre-monsoon and monsoon periods in urban and rural areas of Thiruvananthapuram district. Methods: Based on the daily reports of dengue cases, 70 clusters were identified in Thiruvananthapuram district. A cross-sectional larval survey was done in the domestic and peri-domestic areas of 1750 houses, using the WHO standard techniques. The larval indices were calculated, and the larvae were identified by using taxonomic keys. Urban and rural differences and the variations during pre-monsoon and monsoon seasons were also studied. Results: In the surveyed houses, 15% had mosquito breeding, with 88% having Aedes larvae. The house index, container index and the breteau index were 13.08, 13.28 and 16.57%, respectively. About 86% of the clusters were found positive for Aedes albopictus and 11% for Ae. aegypti. Aedes albopictus was distributed almost equally in rural and urban clusters, whereas the distribution of Ae. aegypti was significantly higher in urban areas (p = 0.03. The most common water holding containers found (outdoor were of plastic, followed by coconut shells. The breeding preference ratio was highest for tyres. Significantly lesser positivity was found for containers during monsoon period when compared to pre-monsoon period. Conclusion: The geographical distribution of Ae. albopictus is significantly high in peri-domestic areas and, therefore, its epidemiological role in the widespread disease occurrence needs to be studied. The discarded tyres being the most preferred breeding sites, where IEC activities will help in source reduction.

Full Text Available Consequences of larval competition at the population level provide explanation for the differences in relative abundance of Aedes aegypti and Aedes albopictus in different geographical regions. The outcome of competition is assessed through the estimates of the life history traits as a response to varying density and resource available for larval development. In the present study, variations in the life history traits due to density-dependent intra- and inter- specific competition involving A. aegypti and A. albopictus were assessed following the minimalist model. The instar-I larvae (0-day old F2 generation of both Aedes species were reared to the adult stages using the initial rearing density of 1, 2, 4 and 6 (individuals/10ml in multiple replicates. The age at pupation, pupal weight, adult weight and adult wing length of the individuals were considered as the response variables and surrogates of estimating the competitive interactions. Density dependent variations in the competitive interactions were evident for both the mosquitoes with reference to the selected life history traits. In A. aegypti, the life history traits varied with the levels of competition, which was not observed for A. albopictus. Although the density levels considered in the present instance were lower than in earlier studies, the observations were similar, with A. albopictus being competitively superior. It appears that irrespective of the density levels, interspecific competition affects A. aegypti and thus may bear population level consequences and overall abundance in the areas where both species are present.

The eggs of the mosquitoAedes aegypti possess the ability to undergo an extended quiescence hosting a fully developed 1st instar larvae within the chorion. As a result of this life history trait pharate larvae can withstand months of quiescence inside the egg where they depend on stored maternal reserves. A. aegypti mosquitoes are frequently associated with urban habitats that may contain significant metal pollution. Therefore, the duration of quiescence and extent of nutritional depletion may affect the physiology and survival of larvae that hatch in a suboptimal habitat. The aim of this study was to determine the effect of an extended quiescence on larval nutrient reserves and the subsequent effects of metal exposure on larval fitness, survival and development. We hypothesized that an extended quiescence would reduce nutritional reserves and alter the molecular response to metal exposure thereby reducing larval survival and altering larval development. As a molecular marker for metal stress responses, we evaluated transcriptional changes in the metallothionein gene (AaMtn) in response to quiescence and metal exposure. Extended 1st instar quiescence resulted in a significant decrease in lipid reserves and negatively affected larval fitness and development. AaMtn transcription and metal tolerance were compromised in first instars emerged from eggs that had undergone an extended quiescence. These findings suggest that newly emerged mosquito larvae that had survived a relatively long pharate 1st instar quiescence (as might occur during a dry season) are more vulnerable to environmental stress. Pharate 1st instar quiescence could have implications for vector control strategies. Newly emerged mosquito larvae at the end of the dry season or start of the wet season are physiologically compromised, and therefore potentially more susceptible to vector control strategies than mosquito larvae hatched subsequently throughout the wet season. Published by Elsevier Ltd.

Adjacent to the northern suburbs of Darwin is a coastal wetland that contains important larval habitats for Aedes vigilax (Skuse), the northern salt marsh mosquito. This species is a vector for Ross River virus and Barmah Forest virus, as well as an appreciable human pest. In order to improve aerial larval control efforts, we sought to identify the most important vegetation categories and climatic/seasonal aspects associated with control operations in these wetlands. By using a generalized linear model to compare aerial control for each vegetation category, we found that Schoenoplectus/mangrove areas require the greatest amount of control for tide-only events (30.1%), and also extensive control for tide and rain events coinciding (18.2%). Our results further indicate that tide-affected reticulate vegetation indicated by the marsh grasses Sporobolus virginicus and Xerochloa imberbis require extensive control for Ae. vigilax larvae after rain-only events (44.7%), and tide and rain events coinciding (38.0%). The analyses of vector control efforts by month indicated that September to January, with a peak in November and December, required the most control. A companion paper identifies the vegetation categories most associated with Aedes vigilax larvae population densities in the coastal wetland. To maximize the efficiency of aerial salt marsh mosquito control operations in northern Australia, aerial control efforts should concentrate on the vegetation categories with high larval densities between September and January.

Malaria is an important cause of illness and death in people living in many parts of the world, especially sub-Saharan Africa. Long-lasting insecticide treated bed nets (LLINs) and indoor residual spraying (IRS) reduce malaria transmission by targeting the adult mosquito vector and are key components of malaria control programmes. However, mosquito numbers may also be reduced by larval source management (LSM), which targets mosquito larvae as they mature in aquatic habitats. This is conducted by permanently or temporarily reducing the availability of larval habitats (habitat modification and habitat manipulation), or by adding substances to standing water that either kill or inhibit the development of larvae (larviciding). To evaluate the effectiveness of mosquito LSM for preventing malaria. We searched the Cochrane Infectious Diseases Group Specialized Register; Cochrane Central Register of Controlled Trials (CENTRAL); MEDLINE; EMBASE; CABS Abstracts; and LILACS up to 24 October 2012. We handsearched the Tropical Diseases Bulletin from 1900 to 2010, the archives of the World Health Organization (up to 11 February 2011), and the literature database of the Armed Forces Pest Management Board (up to 2 March 2011). We also contacted colleagues in the field for relevant articles. We included cluster randomized controlled trials (cluster-RCTs), controlled before-and-after trials with at least one year of baseline data, and randomized cross-over trials that compared LSM with no LSM for malaria control. We excluded trials that evaluated biological control of anopheline mosquitoes with larvivorous fish. At least two authors assessed each trial for eligibility. We extracted data and at least two authors independently determined the risk of bias in the included studies. We resolved all disagreements through discussion with a third author. We analyzed the data using Review Manager 5 software. We included 13 studies; four cluster-RCTs, eight controlled before

Background Malaria is an important cause of illness and death in people living in many parts of the world, especially sub-Saharan Africa. Long-lasting insecticide treated bed nets (LLINs) and indoor residual spraying (IRS) reduce malaria transmission by targeting the adult mosquito vector and are key components of malaria control programmes. However, mosquito numbers may also be reduced by larval source management (LSM), which targets mosquito larvae as they mature in aquatic habitats. This is conducted by permanently or temporarily reducing the availability of larval habitats (habitat modification and habitat manipulation), or by adding substances to standing water that either kill or inhibit the development of larvae (larviciding). Objectives To evaluate the effectiveness of mosquito LSM for preventing malaria. Search methods We searched the Cochrane Infectious Diseases Group Specialized Register; Cochrane Central Register of Controlled Trials (CENTRAL); MEDLINE; EMBASE; CABS Abstracts; and LILACS up to 24 October 2012. We handsearched the Tropical Diseases Bulletin from 1900 to 2010, the archives of the World Health Organization (up to 11 February 2011), and the literature database of the Armed Forces Pest Management Board (up to 2 March 2011). We also contacted colleagues in the field for relevant articles. Selection criteria We included cluster randomized controlled trials (cluster-RCTs), controlled before-and-after trials with at least one year of baseline data, and randomized cross-over trials that compared LSM with no LSM for malaria control. We excluded trials that evaluated biological control of anopheline mosquitoes with larvivorous fish. Data collection and analysis At least two authors assessed each trial for eligibility. We extracted data and at least two authors independently determined the risk of bias in the included studies. We resolved all disagreements through discussion with a third author. We analyzed the data using Review Manager 5 software

Environmental factors such as temperature, nutrient availability, and larval density determine the outcome of postembryonic development in mosquitoes. Suboptimal temperatures, crowding, and starvation during the larval phase reduce adult mosquito size, nutrient stores and affect vectorial capacity. In this study we compared adult female Aedes aegypti, Rockefeller strain, raised under standard laboratory conditions (Large) with those raised under crowded and nutritionally deprived conditions (Small). To compare the gene expression and nutritional state of the major energy storage and metabolic organ, the fat body, we performed transcriptomics using Illumina based RNA-seq and metabolomics using GC/MS on females before and 24 hours following blood feeding. Analysis of fat body gene expression between the experimental groups revealed a large number of significantly differentially expressed genes. Transcripts related to immunity, reproduction, autophagy, several metabolic pathways; including amino acid degradation and metabolism; and membrane transport were differentially expressed. Metabolite profiling identified 60 metabolites within the fat body to be significantly affected between small and large mosquitoes, with the majority of detected free amino acids at a higher level in small mosquitoes compared to large. Gene expression and metabolites in the adult fat body reflect the individual post-embryonic developmental history of a mosquito larva. These changes affect nutritional storage and utilization, immunity, and reproduction. Therefore, it is apparent that changes in larval environment due to weather conditions, nutrition availability, vector control efforts, and other factors can affect adult vectorial capacity in the field.

Introduction of irrigation projects in developing nations has often been blamed for aggravating the problem of mosquito-borne diseases by creating ideal larval habitats for vector mosquitoes. However, whereas several studies have demonstrated the relationship between malaria vectors and irrigation, little work has been done on culicine mosquitoes despite their potential in transmission of filariasis and arboviruses and their significant biting nuisance in these areas. This study examined the diversity of Culex mosquito fauna and their larval habitats at two sites (Murinduko and Kiamachiri) in Mwea, Kenya over a 12-month period. The habitat types present at each site within a 200-meter radius around the study village, including randomly selected paddies and canals, were sampled every two weeks to examine the relationship between vegetation cover, water depth, turbidity, and Culex larval counts. Ten culicine species belonging to four genera were identified, with 73.1% of the total collection comprising of Culex duttoni and Cx. quinquefasciatus. Other species collected included Cx. annulioris, Cx. poicilipes, Cx. cinereus, Cx. tigripes, Cx. trifilatus, Aedes spp., Coquilettidia fuscopennata, and Ficalbia splendens. Murinduko was more diverse than Kiamachiri in terms of species richness (10 versus 7 species) and larval habitat diversity (11 versus 8 habitat types). Paddies, canals, and rain pools were the most diverse habitats in terms of species richness, and ditches, rock pools, and tree holes were the least diverse. Principal component and correlation analyses showed a strong association between three Culex species and the measured habitat characteristics. Culex poicilipes was strongly associated with floating vegetation, Cx. annulioris with clean water containing emergent vegetation, and Cx. quinquefasciatus was associated with turbid water. Seasonal changes in larval counts in water reservoirs and pool and ditch habitats were closely associated with rainfall

Allochthonous leaf litter is often the main resource base for invertebrate communities in ephemeral water-filled containers, and detritus quality can be affected by hydrologic conditions. The invasive mosquitoAedes albopictus utilizes container habitats for its development where it competes as larvae for detritus and associated microorganisms with the native Aedes triseriatus. Different hydrologic conditions that containers are exposed to prior to mosquito utilization affect litter decay and associated water quality. We tested the hypothesis that larval competition between A. albopictus and A. triseriatus would be differentially affected by prior hydrologic conditions. Experimental microcosms provisioned with Quercus alba L. litter were subjected to one of three different hydrologic treatments prior to the addition of water and mosquito larvae: dry, flooded, and a wet/dry cycle. Interspecific competition between A. albopictus and A. triseriatus was mediated by hydrologic treatment, and was strongest in the dry treatment vs. the flooded or wet/dry treatments. Aedes triseriatus estimated rate of population change (λ') was lowest in the dry treatment. Aedes albopictus λ' was unaffected by hydrologic treatment, and was on average always increasing (i.e., > 1). Aedes triseriatus λ' was affected by the interaction of hydrologic treatment with interspecific competition, and was on average declining (i.e., litter had the slowest decay rate and leached the highest concentration of tannin-lignin, but supported more total bacteria than the other treatments. These results suggest that dry conditions negatively impact A. triseriatus population performance and may result in the competitive exclusion of A. triseriatus by A. albopictus, possibly by reducing microbial taxa that Aedes species browse. Changing rainfall patterns with climate change are likely to affect competition between A. triseriatus and A. albopictus, probably enhancing negative competitive effects of A

Natural insecticides from plant origin against mosquito vectors have been the main concern for research due to their high level of eco-safety. Control of mosquitoes in their larval stages are an ideal method since Aedes larvae are aquatic, thus it is easier to deal with them in this habitat. The present study was specifically conducted to explore the larvicidal efficacy of different plant parts of Ipomoea cairica (L.) or railway creeper crude extract obtained using two different solvents; methanol and acetone against late third-stage larvae of Aedes albopictus (Skuse) and Aedes aegypti (L.) (Diptera: Culicidae). Plant materials of I. cairica leaf, flower, and stem were segregated, airdried, powdered, and extracted using Soxhlet apparatus. Larvicidal bioassays were performed by using World Health Organization standard larval susceptibility test method for each species which were conducted separately for different concentration ranging from 10 to 450 ppm. Both acetone and methanol extracts showed 100% mortality at highest concentration tested (450 ppm) after 24 h of exposure. Results from factorial ANOVA indicated that there were significant differences in larvicidal effects between mosquito species, solvent used and plant parts (F=5.71, df=2, Paegypti with LC50 of 101.94 ppm followed by Ae. albopictus with LC50 of 105.59 ppm compared with other fractions of I. cairica extract obtained from flower, stem, and when methanol are used as solvent. The larvae of Ae. aegypti appeared to be more susceptible to I. cairica extract with lower LC50 value compared with Ae. albopictus (F=8.83, df=1, Pinsecticide for the control of Aedesmosquitoes. This study quantified the larvicidal property of I. cairica extract, providing information on lethal concentration that may have potential for a more eco-friendly Aedesmosquito control program.

A cDNA clone originating from adult female Aedes aegypti mosquitoes was found with substantial similarity to nucleosidases of the EC 3.2.2.1 enzyme class. Although this type of enzyme is unusual in animals, abundant enzyme activity was found in salivary homogenates of this mosquito, but not in salivary homogenates of the mosquitoes Anopheles gambiae and Culex quinquefasciatus, or the sand fly Lutzomyia longipalpis. Aedes salivary homogenate hydrolyses inosine and guanosine to hypoxanthine and xanthine plus the ribose moiety, but does not hydrolyse the pyrimidines uridine and cytidine, thus characterizing the presence of a purine nucleosidase activity. The enzyme is present in oil-induced saliva, indicating that it is secreted. Male Ae. aegypti salivary gland homogenates (SGH) have very low purine nucleosidase activity, suggesting that the enzyme plays a role in mosquito blood feeding. A novel isocratic HPLC method to separate nucleosides and their bases is described.

This study attempted to find organisms for the biological control of the mosquitoAedes aegypti in Costa Rica. Copepods of the genera Arctodiaptomus, Eucylops, Mesocyclops, Megacyclops, and Thermocyclops were collected in several parts of the country and cultured for laboratory evaluations. Mesocyclops thermocyclopoides was the most successful species in reducing the number of larval Ae. aegypti (7.3 larvae in 24 h at a density of 200 Aedes/liter). Arctodiaptomus dorsalis, Eucyclops cf. bondi, Eucyclops leptacanthus, Megacyclops sp., and Thermocyclops decipens were not effective predators. In cage simulation trials, M. thermocyclopoides showed 100% larval reduction after 4 wk and adult mosquitoes disappeared after 7 wk. The copepod was able to survive in Aechmea sp. bromeliads under laboratory conditions. In field trials under 3 different climatic conditions M. thermocyclopoides survived 2-5 months in bromeliad leaf axils and 3-6 months in used car tires. In tires, this species reduced the number of larval Ae. aegypti 79, 90, and 99% in tropical dry, moderate, and humid climates, respectively. An El Niño phenomenon affected the results by drought, which apparently also caused a decline in the population of the predatory mosquito Toxorhynchites haemorrhoidalis superbus. Considering these severe test conditions, M. thermocyclopoides might be a promising predator for mosquito control in Costa Rica.

Full Text Available An important question for mosquito population dynamics, mosquito-borne pathogen transmission and vector control is how mosquito populations are regulated. Here we develop simple models with heterogeneity in egg laying patterns and in the responses of larval populations to crowding in aquatic habitats. We use the models to evaluate how such heterogeneity affects mosquito population regulation and the effects of larval source management (LSM. We revisit the notion of a carrying capacity and show how heterogeneity changes our understanding of density dependence and the outcome of LSM. Crowding in and productivity of aquatic habitats is highly uneven unless egg-laying distributions are fine-tuned to match the distribution of habitats' carrying capacities. LSM reduces mosquito population density linearly with coverage if adult mosquitoes avoid laying eggs in treated habitats, but quadratically if eggs are laid in treated habitats and the effort is therefore wasted (i.e., treating 50% of habitat reduces mosquito density by approximately 75%. Unsurprisingly, targeting (i.e. treating a subset of the most productive pools gives much larger reductions for similar coverage, but with poor targeting, increasing coverage could increase adult mosquito population densities if eggs are laid in higher capacity habitats. Our analysis suggests that, in some contexts, LSM models that accounts for heterogeneity in production of adult mosquitoes provide theoretical support for pursuing mosquito-borne disease prevention through strategic and repeated application of modern larvicides.

Full Text Available Abstract Background Genetic modification of mosquitoes offers a promising strategy for the prevention and control of mosquito-borne diseases. For such a strategy to be effective, it is critically important that engineered strains are competitive enough to serve their intended function in population replacement or reduction of wild mosquitoes in nature. Thus far, fitness evaluations of genetically modified strains have not addressed the effects of competition among the aquatic stages and its consequences for adult fitness. We therefore tested the competitive success of combinations of wild, inbred and transgenic (created in the inbred background immature stages of the dengue vector Aedes aegypti in the presence of optimal and sub-optimal larval diets. Results The wild strain of Ae. aegypti demonstrated greater performance (based on a composite index of survival, development rate and size than the inbred strain, which in turn demonstrated greater performance than the genetically modified strain. Moreover, increasing competition through lowering the amount of diet available per larva affected fitness disproportionately: transgenic larvae had a reduced index of performance (95-119% compared to inbred (50-88% and wild type larvae (38-54%. In terms of teneral energy reserves (glycogen, lipid and sugar, adult wild type mosquitoes had more reserves directly available for flight, dispersal and basic metabolic functions than transgenic and inbred mosquitoes. Conclusions Our study provides a detailed assessment of inter- and intra-strain competition across aquatic stages of wild type, inbred, and transgenic mosquitoes and the impact of these conditions on adult energy reserves. Although it is not clear what competitive level is adequate for success of transgenic strains in nature, strong gene drive mechanisms are likely to be necessary in order to overcome competitive disadvantages in the larval stage that carryover to affect adult fitness.

Larvicides target larvae in the breeding habitat before they can mature into adult mosquitoes and disperse. Larvicide treatment of breeding habitats help reduce the adult mosquito population in nearby areas.

Full Text Available Mosquitoes are well known as vectors of several disease causing pathogens. The extensive use of synthetic insecticides in the mosquito control strategies resulted to the development of pesticide resistance and fostered environmental deterioration. Hence in recent years plants become alternative source of mosquito control agents. The present study assessed the larvicidal and oviposition altering activity of six different plants species-Alstonia scholaris, Callistemon viminalis, Hyptis suaveolens, Malvastrum coromandelianum, Prosopis juliflora, Vernonia cinerea against Aedes albopictus mosquito in laboratory.Leaf extracts of all the six plants species in five different solvents of various polarities were used in the range of 20-400ppm for larval bioassay and 50,100 and 200ppm for cage bioassay (for the study of oviposition behavior against Ae. albopictus. The larval mortality data were recorded after 24 h and subjected to Probit analysis to determine the lethal concentrations (LC50, while OAI (Oviposition activity index was calculated for oviposition altering activity of the plant extracts.Vernonia cinerea extract in acetone and C. viminalis extract in isopropanol were highly effective against Aedes albopictus larvae with LC50 value 64.57, 71.34ppm respectively. Acetone extract of P. juliflora found to be strong oviposition-deterrent which inhibited >2 fold egg laying (OAI-0.466 at 100ppm.Vernonia cinerea and C. viminallis leaf extracts have the potential to be used as larvicide and P. juliflora as an oviposition-deterrent for the control of Ae. albopictus mosquito.

As evidence linking Zika virus with serious health complications strengthens, public health officials and clinicians worldwide need to know which locations are likely to be at risk for autochthonous Zika infections. We created risk maps for epidemic and endemic Aedes-borne Zika virus infections globally using a predictive analysis method that draws on temperature, precipitation, elevation, land cover, and population density variables to identify locations suitable for mosquito activity seasonally or year-round. Aedesmosquitoes capable of transmitting Zika and other viruses are likely to live year-round across many tropical areas in the Americas, Africa, and Asia. Our map provides an enhanced global projection of where vector control initiatives may be most valuable for reducing the risk of Zika virus and other Aedes-borne infections.

Mosquito populations are largely regulated by processes occurring at the larval stage. We sampled mosquitolarval microhabitats (mostly water-holding containers) in six neighborhoods in the Washington, DC, area that varied in socioeconomic status (SES) and housing structure (row houses vs. stand-alone houses) to test associations among these neighborhood characteristics, microhabitat abundance and parameters, and mosquito occurrence and densities. Thirty-four percent (33.9%) of sampled microhabitats contained mosquito larvae, and 93.1% of larvae were Aedes albopictus Skuse or Culex pipiens L. Five specific container types (drains, corrugated flexible drainpipes, planters, garbage cans, and buckets) accounted for the majority of water-holding (56.0%) and mosquito-positive (50.6%) microhabitats sampled. We found no associations between SES or housing structure with total microhabitat abundance per yard, mosquito occurrence or mosquito densities per microhabitat. In contrast, container purpose varied with SES, with low SES neighborhoods having greater numbers of disused containers and lower numbers of functional containers than low and medium SES neighborhoods. Ae. albopictus were 83% more abundant in disused containers, whereas Cx. pipiens were more abundant in structural and functional containers, possibly owing to species-specific oviposition and development related to water quality. Ae. albopictus densities increased over the summer, whereas Cx. pipiens densities remained constant. Ae. albopictus is usually the dominant pest in urban areas in the eastern United States; therefore, integrated mosquito management programs should incorporate the elimination of disused containers to reduce its infestation and adult production, especially in low SES neighborhoods where they occur most frequently.

To test whether Zika virus has adapted for more efficient transmission by Aedes aegypti mosquitoes, leading to recent urban outbreaks, we fed mosquitoes from Brazil, the Dominican Republic, and the United States artificial blood meals containing 1 of 3 Zika virus strains (Senegal, Cambodia, Mexico) and monitored infection, dissemination, and virus in saliva. Contrary to our hypothesis, Cambodia and Mexica strains were less infectious than the Senegal strain. Only mosquitoes from the Dominican Republic transmitted the Cambodia and Mexica strains. However, blood meals from viremic mice were more infectious than artificial blood meals of comparable doses; the Cambodia strain was not transmitted by mosquitoes from Brazil after artificial blood meals, whereas 61% transmission occurred after a murine blood meal (saliva titers up to 4 log10 infectious units/collection). Although regional origins of vector populations and virus strain influence transmission efficiency, Ae. aegypti mosquitoes appear to be competent vectors of Zika virus in several regions of the Americas. PMID:28287375

To test whether Zika virus has adapted for more efficient transmission by Aedes aegypti mosquitoes, leading to recent urban outbreaks, we fed mosquitoes from Brazil, the Dominican Republic, and the United States artificial blood meals containing 1 of 3 Zika virus strains (Senegal, Cambodia, Mexico) and monitored infection, dissemination, and virus in saliva. Contrary to our hypothesis, Cambodia and Mexica strains were less infectious than the Senegal strain. Only mosquitoes from the Dominican Republic transmitted the Cambodia and Mexica strains. However, blood meals from viremic mice were more infectious than artificial blood meals of comparable doses; the Cambodia strain was not transmitted by mosquitoes from Brazil after artificial blood meals, whereas 61% transmission occurred after a murine blood meal (saliva titers up to 4 log 10 infectious units/collection). Although regional origins of vector populations and virus strain influence transmission efficiency, Ae. aegypti mosquitoes appear to be competent vectors of Zika virus in several regions of the Americas.

RNA interference (RNAi) is a powerful approach for elucidating gene functions in a variety of organisms, including mosquitoes and many other insects. Little has been done, however, to harness this approach in order to control adult and larvalmosquitoes. Juvenile hormone (JH) plays a pivotal role in the control of reproduction in adults and metamorphism in larvalmosquitoes. This report describes an approach to control Aedes aegypti using RNAi against JH acid methyl transferase (AeaJHAMT), the ultimate enzyme in the biosynthetic pathway of JH III that converts JH acid III (JHA III) into JH III. In female A. aegypti that were injected or fed jmtA dsRNA targeting the AeaJHAMT gene (jmtA) transcript, egg development was inhibited in 50% of the treated females. In mosquito larvae that were fed transgenic Pichia pastoris cells expressing long hair pin (LHP) RNA, adult eclosion was delayed by 3 weeks causing high mortality. Northern blot analyses and qPCR studies show that jmtA dsRNA causes inhibition of jmtA transcript in adults and larvae, which is consistent with the observed inhibition of egg maturation and larval development. Taken together, these results suggest that jmtA LHP RNA expressed in heat inactivated genetically modified P. pastoris cells could be used to control mosquito populations in the marsh.

Insecticide applications are not particularly effective on Aedes aegypti mosquitoes which has been attributed to their 'closet' behaviour, or ability to rest in places that remain unexposed to insecticides. Some researchers have suggested that insecticides repel mosquitoes, which would result in less exposure and increased dispersal. If repellence due to insecticides is a fact, acquiring a vector-borne disease, such as dengue, could legitimately be attributed to local vector control efforts and this would lead to restitution claims. This study thus investigated the effect of insecticide presence on mosquito behaviour indirectly via oviposition and directly via olfactory response. In all experiments, oviposition in each insecticide compared to its water and ethanol controls was not significantly different. This indicates that Ae. aegypti mosquitoes are not affected by insecticide presence and that increased dispersal is unlikely to be caused by vector control spraying.

Full Text Available The mosquitoAedes albopictus, indigenous to Southeast Asia and nearby islands, has spread almost worldwide during recent decades. We confirm the invasion of this mosquito, first reported in Yamagata city in northeast Honshu, Japan in 2000. Previously, only Ae. japonicus japonicus had been collected in this place, but 2 years later, the population of Ae. albopictus had increased, so more than 80% of the total number of larval colonies there consisted of this species. In contrast to Yamagata’s new residential area, now infested by Ae. albopictus, the original mosquito remains in the city but its habitats are generally closer to the surrounding mountains, where the normalized difference vegetation index is higher. The factors affecting the distribution of both species in Yamagata city were studied using geographical information systems (GIS based on data derived from field surveys, aerial photographs, satellite images and digital maps. The range of Aedesmosquito habitats was estimated and visualised on polygon maps and no significant differences were noted when the polygon area was calculated by GIS software in comparison with the satellite images. Although Ae. j. japonicus was expected to be rapidly overrun by Ae. albopictus, this did not happen. Currently, both species coexist; not only in separate sites, but also simultaneously in various water bodies, where larvae from both species have frequently been seen. However, the competitive relationship between these two Aedes species within a warming environment is an issue that should be closely monitored.

Consequences of larval competition at the population level provide explanation for the differences in relative abundance of Aedes aegypti and Aedes albopictus in different geographical regions. The outcome of competition is assessed through the estimates of the life history traits as a response to varying density and resource available for larval development. In the present study, variations in the life history traits due to density-dependent intra- and inter- specific competition involving A...

The efficacy of maximum label rates of bifenthrin applications to dry tires to prevent Aedesmosquito breeding was investigated by field colonization and bioassay trials in shaded and unshaded locations. Aedes notoscriptus and Culex quinquefasciatus larvae were the most abundant species present in the field colonization trial. Colonization and survival of Ae. notoscriptus larvae to the late instar occurred significantly earlier in treated tires in shaded compared with unshaded locations (P = 0.002). Bifenthrin applications in shaded tires only prevented early instar survival for approximately 2.6 wk. Aedes notoscriptus late instars did not appear in the treated unshaded tires. Culex quinquefasciatus colonized treated tires from the 2nd wk in both shaded and unshaded treatments. In the bioassay, water from bifenthrin-treated tires, through extrapolation, was found to kill approximately 100% of late instar Ae. notoscriptus for only approximately 2.0-2.2 wk in shaded and unshaded tires. Under conditions optimal for Aedes breeding, such as shaded locations, high ambient temperatures, high relative humidity, and high amounts of leaf/organic matter accumulations, bifenthrin may not be effective as a larval control measure in tires for greater than 2.0-2.6 wk.

Full Text Available Aedes (Stegomyia albopictus (Skuse and Aedes (Stegomyia cretinus Edwards are closely related mosquito species with common morphological features and bio-ecological similarities. Recent mosquito surveillance in Athens, Greece, showed that they are sympatric mosquito species, with Ae. albopictus [corrected] developing quite higher population densities than Ae. cretinus [corrected] . The potential of mating interference between these species was investigated by reciprocal and homologous mating experiments in cages under laboratory conditions. In non-choice interspecific crosses (groups of males and females females of both species produced sterile eggs. Insemination rate was 58% for Ae. cretinus females and only 1% for Ae. albopictus [corrected] females. Aedes albopictus males were sexually aggressive and inseminated Ae. cretinus females (31% in choice experiments, where males of one species had access to mate with females of both species. Whereas, interspecific mating of Ae. albopictus [corrected] females with Ae. cretinus males in the co-occurrence of Ae. cretinus females was weaker (4%. Aedes cretinus females from non-choice crossing with Ae. albopictus [corrected] or Ae. cretinus males were paired individually with conspecific males. The percentage of fertile Ae. cretinus females was 17.5% when had encaged before with Ae. albopictus [corrected] males, compared to 100% when Ae. cretinus [corrected] females were encaged with conspecific males only. Probable ecological consequences of asymmetric mating between these ecologically homologous species in nature are discussed.

ABSTRACT Certain strains of the intracellular endosymbiont Wolbachia can strongly inhibit or block the transmission of viruses such as dengue virus (DENV) by Aedesmosquitoes, and the mechanisms responsible are still not well understood. Direct infusion and liquid chromatography-Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometry-based lipidomics analyses were conducted using Aedes albopictus Aa23 cells that were infected with the wMel and wMelPop strains of Wolbachia in comparison to uninfected Aa23-T cells. Substantial shifts in the cellular lipid profile were apparent in the presence of Wolbachia. Most significantly, almost all sphingolipid classes were depleted, and some reductions in diacylglycerols and phosphatidylcholines were also observed. These lipid classes have previously been shown to be selectively enriched in DENV-infected mosquito cells, suggesting that Wolbachia may produce a cellular lipid environment that is antagonistic to viral replication. The data improve our understanding of the intracellular interactions between Wolbachia and mosquitoes. IMPORTANCE Mosquitoes transmit a variety of important viruses to humans, such as dengue virus and Zika virus. Certain strains of the intracellular bacterial genus called Wolbachia found in or introduced into mosquitoes can block the transmission of viruses, including dengue virus, but the mechanisms responsible are not well understood. We found substantial shifts in the cellular lipid profiles in the presence of these bacteria. Some lipid classes previously shown to be enriched in dengue virus-infected mosquito cells were depleted in the presence of Wolbachia, suggesting that Wolbachia may produce a cellular lipid environment that inhibits mosquito-borne viruses. PMID:26994075

Aedes aegypti is the main insect vector of Dengue fever and dengue hemorrhagic fever/dengue shock syndrome and represents the only vulnerable element in the control of this disease. Therefore, the identification and quantification of this mosquito is an important task; however, the majority of taxonomic keys are based on the 4th larval instar. For that reason, this study describes the four larval instars ofA. aegypti using scanning electron microscopy. Morphological changes during larval development were observed at the pecten, comb scales and the ventral brush of the abdominal segment X; however, the 3rd and 4th instars showed similar structures with only a slight variation. The structures described in this study will be helpful in the identification of the four instars of A. aegypti, a fundamental task for comprehending the natural history of dengue mainly in new territories affected.

We evaluated degree-day predictions of Dirofilaria immitis development (HDU) under constant and fluctuating temperature treatments of equal average daily temperature. Aedes aegypti mosquitoes were infected with D. immitis microfilariae and parasite development was recorded at set time points in dissected mosquitoes. Time to L3 development in Malpighian tubules and detection in mosquito heads was shorter for larvae experiencing a daily regime of 19±9°C than larvae at constant 19°C; larval development rate in Malpighian tubules was slower in fluctuating regimes maintained above the 14°C developmental threshold than larvae under constant temperatures. We showed that hourly temperature modeling more accurately predicted D. immitis development to infective L3 stage. Development time differed between fluctuating and constant temperature treatments spanning the 14°C development threshold, implicating a physiological basis for these discrepancies. We conclude that average daily temperature models underestimate L3 development—and consequently dog heartworm transmission risk—at colder temperatures, and spatio-temporal models of D. immitis transmission risk should use hourly temperature data when analyzing high daily temperature ranges spanning 14°C. PMID:25747489

Monitoring dengue vector control by sampling adult Aedes aegypti (L.) recently has been used to replace both larval and pupal surveys. We have developed and evaluated the Gravid Aedes Trap (GAT) through a sequential behavioral study. The GAT does not require electricity to function, and trapped mosquitoes are identified easily during trap inspections. The GAT concept relies on visual and olfactory cues to lure gravid Ae. aegypti and an insecticide to kill trapped mosquitoes. Gravid mosquitoes are lured to a black bucket base containing oviposition attractant (infusion) and are trapped in a translucent chamber impregnated with a pyrethroid insecticide where they are killed within 3-15 min. In semifield observations, the GAT captured a significantly higher proportion of gravid mosquitoes than the double sticky ovitrap. We also demonstrated that the visual cues of the prototype GAT-LgBF (large black base bucket with a black funnel at the top of the translucent chamber) captured a significantly higher proportion of gravid mosquitoes than the other prototypes. The visual contrast created by the addition of a white lid to the top of the black funnel significantly increased the number of captured gravid mosquitoes when compared with the GAT-LgBF in semifield trials. We conclude that the GAT is more efficient in recapturing gravid Ae. aegypti when compared with sticky ovitraps. The GAT is an effective, practical, low cost, and easily transportable trap, features that are essential in large-scale monitoring programs, particularly in areas where funding is limited.

To investigate whether the unique assemblage of habitats in zoos could affect mosquito oviposition behavior and to provide zoos with suggestions for mosquito control, larvae were sampled and associated habitat variables were measured in 2 zoos in South Carolina, U.S.A. Fifty-nine sites were sampled from March 2008 to January 2009. A total of 1630 larvae representing 16 species was collected and identified. The dominant species was Aedes albopictus (46.0%), followed by Ae. triseriatus (23.6%), Culex restuans (12.4%), and Cx. pipiens complex (9.7%). Principal components and multiple logistic regression analyses showed that across both zoos the distribution of Ae. albopictus larvae was predicted by ambient and site temperature, precipitation, dissolved oxygen, and container habitats. The distribution of Ae. triseriatus larvae was predicted by natural containers and shade height zoo personnel include elimination of artificial container habitats, reduction of shade sources < or =2 m over aquatic habitats, use of approved mosquito larvicides, and training in recognizing and mitigating larvalmosquito habitats.

Few studies have taken a comprehensive approach of measuring the impact of inter- and intra-specific larval competition on adult mosquito traits. In this study, the impact of competition Aedes aegypti and A. albopictus was quantified over the entire life of a cohort.Competitive treatments affected hatch-to-adult survivorship and development time to adulthood of females for both species, but affected median wing length of females only for A. albopictus. Competitive treatments had no significant effect on the median adult female longevity nor were there any effects on other individual traits related to bloodfeeding and reproductive success.Analysis of life table traits revealed no effect of competitive treatment on net reproductive rate (R0) but there were significant effects on cohort generation time (Tc) and cohort rate of increase (r) for both species.Inter-specific and intra-specific competition among Aedes larvae may produce individual and population-level effects that are manifest in adults; however, benign conditions may enable resulting adults to compensate for some impacts of competition, particularly those affecting blood feeding success, fecundity, and net reproductive rate, R0. The effect of competition, therefore, affects primarily larva - to - adult survivorship and larval development time, which in turn impact the cohort generation time, Tc and ultimately cohort rate of increase, r.The lack of effects of larval rearing environment on adult longevity suggests that effects on vectorial capacity due to longevity may be limited if adults have easy access to sugar and blood meals.

Objective: To evaluate the potential of local mosquitoes to act as vectors for dengue transmission in Japan.Methods: Serotype 2 Th NH28/93 was used to test the dengue susceptibility profiles of Aedes flavopictus miyarai(Ae. f. miyarai), Aedes galloisi(Ae. galloisi) and Aedes albopictus(Ae.albopictus), which were collected in Japan. We used Aedes aegypti from Thailand as a positive control. The mosquitoes were infected with the virus intrathoracically or orally. At 10 or 14 days post infection, the mosquitoes were dissected and total RNA was extracted from their abdomens, thoraxes, heads and legs. Mosquito susceptibility to dengue virus was evaluated using RT-PCR with dengue virus-specific primers. Differences in the infection and mortality rates of the different mosquito species were tested using Fisher’s exact probability test.Results: The infection rates for dengue virus administered intrathoracically to Ae. f. miyarai,Ae. galloisi and Aedes aegypti mosquitoes were identical by RT-PCR on Day 10 post infection.All of the body parts we tested were RT-PCR-positive for dengue virus. For the orally administered virus, the infection rates in the different body parts of the Ae. f. miyarai mosquitoes were slightly higher than those of Ae. albopictus mosquitoes, but were similar to the control mosquitoes(P > 0.05). The mortality rates for Ae. f. miyarai and Ae. albopictus mosquitoes were similar(P = 0.19). Our data indicated that dengue virus was able to replicate and disseminate to secondary infection sites in all of the four mosquito species(Japanese and Thai).Conclusions: Ae. albopictus is a well-known candidate for dengue transmission in Japan. However, our data suggest that Ae. f. miyarai from Ishigaki Island(near Okinawa Island) and Ae. galloisi from Hokkaido(Northern Japan) should also be regarded as potential vectors for dengue transmission in these regions. Further studies on these mosquitoes should be conducted.

Objective: To evaluate the potential of local mosquitoes to act as vectors for dengue transmission in Japan. Methods: Serotype 2 ThNH28/93 was used to test the dengue susceptibility profiles of Aedes flavopictus miyarai (Ae. f. miyarai), Aedes galloisi (Ae. galloisi) and Aedes albopictus (Ae. albopictus), which were collected in Japan. We used Aedes aegypti from Thailand as a positive control. The mosquitoes were infected with the virus intrathoracically or orally. At 10 or 14 days post infection, the mosquitoes were dissected and total RNA was extracted from their abdomens, thoraxes, heads and legs. Mosquito susceptibility to dengue virus was evaluated using RT-PCR with dengue virus-specific primers. Differences in the infection and mortality rates of the different mosquito species were tested using Fisher's exact probability test. Results: The infection rates for dengue virus administered intrathoracically to Ae. f. miyarai, Ae. galloisi and Aedes aegypti mosquitoes were identical by RT-PCR on Day 10 post infection. All of the body parts we tested were RT-PCR-positive for dengue virus. For the orally admin-istered virus, the infection rates in the different body parts of the Ae. f. miyarai mosquitoes were slightly higher than those of Ae. albopictus mosquitoes, but were similar to the control mosquitoes (P>0.05). The mortality rates for Ae. f. miyarai and Ae. albopictus mosquitoes were similar (P=0.19). Our data indicated that dengue virus was able to replicate and disseminate to secondary infection sites in all of the four mosquito species (Japanese and Thai). Conclusions: Ae. albopictus is a well-known candidate for dengue transmission in Japan. However, our data suggest that Ae. f. miyarai from Ishigaki Island (near Okinawa Island) and Ae. galloisi from Hokkaido (Northern Japan) should also be regarded as potential vectors for dengue transmission in these regions. Further studies on these mosquitoes should be conducted.

A countrywide field survey of immature mosquitoes was conducted in Moldova with the aim to evaluate the Culicidae species composition in different larval habitats and their distribution in the country. In total, 259 potential larval habitats were sampled in the 53 localities, resulting in 9,456 specimens. Twenty species belonging to the genera Anopheles, Aedes, Culex, Culiseta, and Uranotaenia were collected. Mean species richness in aquatic habitats ranged from 1.00 to 4.00, and, for example, was higher in swamps, flood plains, ditches, and large ground pools and lower in rivers, streams, tree-holes, and containers. Six mosquito species were identified only in a single type of aquatic habitat. Anopheles maculipennis s.l., Culex pipiens pipiens L., and Culex modestus Ficalbi were the most abundant and distributed species representing over 80% of the identified specimens. Three, four, and five associated species were recorded from 23.5% of mosquito-positive aquatic habitats. Our findings demonstrate the co-occurrence of Cx. p. pipiens and Culex torrentium Martini in natural and rural environments. It is concluded that the study area has undergone a dramatic ecological change since the previous studies in the 1950s, causing the near extinction of Culex theileri Theobald from Moldova. An. maculipennis s.l. larval abundance, reduced by the DDT control of the adults in the 1950s, had returned to those of the 1940s. Restoration of An. maculipennis s.l. abundance in combination with imported malaria cases constitute a risk of the reintroduction of malaria transmission in Moldova.

Full Text Available Background & objectives: The quality of breeding sites is reflected through the pupal productivity and the life history traits of Aedesmosquitoes. Using nutrient reserves and pupal productivity of Aedes as indicators, the larval habitats including sewage drains were characterized to highlight the habitat expansion and vector management. Methods: The pupae and adults collected from the containers and sewage drains were characterized in terms of biomass and nutrient reserves and the data were subjected to three way factorial ANOVA. Discriminant function analyses were performed to highlight the differences among the habitats for sustenance of Aedesmosquitoes. Results: Survey of larval habitats from the study area revealed significant differences (P<0.05 in the pupal productivity of Aedes among the habitats and months. Despite sewage drains being comparatively less utilized for breeding, the pupae were of higher biomass with corresponding adults having longer wings in contrast to other habitats. The nutrient reserve of the adults emerging from pupae of sewage drains was significantly higher (P<0.05, compared to other habitats, as reflected through the discriminant function analysis. Interpretation & conclusions: The present results showed that for both Ae. aegypti and Ae. albopictus, sewage drains were equally congenial habitat as were plastic, porcelain and earthen habitats. Availability of Aedes immature in sewage drains poses increased risk of dengue, and thus vector control programme should consider inclusion of sewage drains as breeding habitat of dengue vector mosquitoes.

Background: Dengue is amongst the most serious mosquito-borne infectious disease with hot spots in tropical and subtropical parts of the world. Unfortunately, no licensed vaccine for the disease is currently available in medicine markets. The only option available is the management of dengue vector mosquito, Aedes aegypti (Diptera: Culicidae). Method: Predatory potential of five odonate nymphs namely Anax parthenope, Bradinopyga geminate, Ischnura forcipata, Rhinocypha quadrimaculata, and Orthetrum sabina were evaluated against the 4th instar larvae of the dengue vector mosquito, Aedes aegypti, under laboratory conditions. The consumption of the mosquito larvae was evaluated at three water volume levels viz., 1 liter, 2 liter and 3 liter. Results: The number of Ae. aegypti larvae consumed varied significantly among the five species, and at different levels of water volume (Podonate nymphs and the water volumes was statistically non-significant (P> 0.05). Ischnura forcipata consumed the highest number of Ae. aegypti larvae (n=56) followed by A. parthenope (n=47) and B. geminate (n=46). The number of larvae consumed was decreased with increasing search area or water volume, and the highest predation was observed at 1-liter water volume. Conclusion: The odonate nymphs could be a good source of biological agents for the management of the mosquitoes at larval stages. PMID:27308283

Full Text Available Background: Dengue is amongst the most serious mosquito-borne infectious disease with hot spots in tropical and subtropical parts of the world. Unfortunately, no licensed vaccine for the disease is currently available in medicine markets. The only option available is the management of dengue vector mosquito, Aedes aegypti (Diptera: Culicidae.Method: Predatory potential of five odonate nymphs namely Anax parthenope, Bradinopyga geminate, Ischnura forcipata, Rhinocypha quadrimaculata, and Orthetrum sabina were evaluated against the 4th instar larvae of the den­gue vector mosquito, Aedes aegypti, under laboratory conditions. The consumption of the mosquito larvae was eval­uated at three water volume levels viz., 1 liter, 2 liter and 3 liter.Results: The number of Ae. aegypti larvae consumed varied significantly among the five species, and at different levels of water volume (P< 0.01. However, the interaction between odonate nymphs and the water volumes was statistically non-significant (P> 0.05. Ischnura forcipata consumed the highest number of Ae. aegypti larvae (n=56 followed by A. parthenope (n=47 and B. geminate (n=46. The number of larvae consumed was decreased with in­creasing search area or water volume, and the highest predation was observed at 1-liter water volume.Conclusion: The odonate nymphs could be a good source of biological agents for the management of the mosquitoes at larval stages.

Sex determination in the mosquitoAedes aegypti is governed by a dominant male-determining factor (M factor) located within a Y chromosome–like region called the M locus. Here, we show that an M-locus gene, Nix, functions as an M factor in A. aegypti. Nix exhibits persistent M linkage and early embryonic expression, two characteristics required of an M factor. Nix knockout with clustered regularly interspaced short palindromic repeats (CRISPR)–Cas9 resulted in largely feminized genetic males ...

SUMMARYParasitic infection is often associated with changes in host life-history traits, such as host development. Many of these life-history changes are ultimately thought to be the result of a depletion or reallocation of the host's resources driven either by the host (to minimize the effects of infection) or by the parasite (to maximize its growth rate). In this paper we investigate the energetic budget of Aedes aegypti mosquito larvae infected by Vavraia culicis, a microsporidian parasite that transmits horizontally between larvae, and which has been previously shown to reduce the probability of pupation of its host. Our results show that infected larvae have significantly less lipids, sugars and glycogen than uninfected larvae. These differences in resources were not due to differences in larval energy intake (feeding rate) or expenditure (metabolic rate). We conclude that the lower energetic resources of infected mosquitoes are the result of the high metabolic demands that microsporidian parasites impose on their hosts. Given the fitness advantages for the parasite of maintaining the host in a larval stage, we discuss whether resource depletion may also be a parasite mechanism to prevent the pupation of the larvae and thus maximize its own transmission.

Objective:To evaluate the growth inhibition activity of the crude extract of Cyperus aromaticus (C. aromaticus) cultured cells against the 3rd instar larvae of Aedes aegypti (Linn.) and Aedes albopictus Skuse (Ae. albopictus) under laboratory conditions, and determine the sublethal effects (EI50) of the crude extract of C. aromaticus cultured cells on some biological and morphological parameters of both Aedesmosquito species during two generations as well. Methods:The cell suspension cultures of C. aromaticus were activated from five callus lines (P4, Pa, Z1, Z6 and Ml) derived from the root explants of in vitro plantlets. The cultured cells were extracted in chloroform and used as plant material for the present study. For detection of juvenile hormone III, the crude extracts were analyzed by HPLC. Then the crude extracts of the three C. aromaticus cultured cell lines which contained varied amounts of juvenile hormone III [high level (P4 cell line), medium level (Z1 cell line) and low level (Ml cell line)] were tested against Aedesmosquito species. Laboratory evaluation was performed against late third instar larvae of the Vector Control Research Unit strains of Ae. aegypti and Ae. albopictus using the standard WHO method. The effects of EI50 of the C. aromaticus cultured P4 cells on fecundity, fertility, growth period, sex ratio, adult size and longevity of Aedesmosquitoes were assessed. Results:Bioassay tests presented the remarkable growth inhibition activity of the crude extracts of C. aromaticus cultured cells against the two Aedesmosquitoes. Between the two mosquito species, Ae. albopictus was more susceptible to the crude extracts with lower EI50 values. EI50 of the crude extract of C. aromaticus cultured cells (P4) increased the sterility indices in the parental generation females in both Aedesmosquito species. A significant delay in the pupal formation and adult emergence were observed in the parental generation of the both mosquito species. The sex

Background Dengue (DENV), Chikungunya (CHIKV), Zika (ZIKV), as well as yellow fever (YFV) viruses are transmitted to humans by Aedes spp. females. In Salvador, the largest urban center in north-eastern Brazil, the four DENV types have been circulating, and more recently, CHIKV and ZIKV have also become common. We studied the role of storm drains as Aedeslarval development and adult resting sites in four neighbourhoods of Salvador, representing different socioeconomic, infrastructure and topo...

Mosquitoes are the most important arthropod disease vectors, transmitting a broad range of pathogens that cause diseases such as malaria, lymphatic filariasis, and yellow fever. Mosquitoes and other insects are able to mount powerful cellular and humoral immune responses against invading pathogens. To date, most studies have concentrated on the humoral response. In the current study we describe the hemocytes (blood cells) of the yellow fever mosquito, Aedes aegypti, by means of morphology, lectin binding, and enzyme activity and immunocytochemistry. Our light and electron microscopic studies suggest the presence of four distinct hemocyte types: granulocytes, oenocytoids, adipohemocytes, and thrombocytoids. We believe granulocytes and oenocytoids are true circulating hemocytes, but adipohemocytes and thrombocytoids are likely adhered to fixed tissues. Granulocytes, the most abundant cell type, have acid phosphatase and alpha-naphthyl acetate esterase activity, and bind the exogenous lectins WGA, HPA, and GNL. Phenoloxidase, an essential enzyme in the melanotic encapsulation immune response, was detected inside oenocytoids. This is, to our knowledge, the first report that has detected phenoloxidase inside mosquito hemocytes at the ultrastructural level. These results have begun to form a knowledge base for our ongoing studies on the function of Ae. aegypti hemocytes, and their involvement in controlling infections.

The mosquitoAedes aegypti is an epidemic vector of several diseases such as dengue fever and yellow fever. Several pesticides are used to control the mosquito population. Because of their frequent use, some mosquitoes have developed resistance. In this study, we used the Y-tube olfactometer to test essential oils of Cymbopogon species and screened specific formulas of components as repellents against Ae. aegypti. At 400 μL, the extracted oil of citronella grass (Cymbopogon nardus) and myrcene produced a low-active response by inhibiting mosquito host-seeking activity. Citronella grass, lemon grass (Cymbopogon citratus), citral and myrcene also produced a low-treatment response to repellents, for more potential to affect host-seeking behavior. Furthermore, the mixture of citral, myrcene, and citronellal oil (C:M:Ci = 6:4:1) greatly affected and inhibited host-seeking behavior (76% active response; 26% treatment response with 40 μL; 42.5%, 18% with 400 μL; and 19%, 23% with 1000 μL). As compared with the result for N,N-diethyl-3-methylbenzamide (DEET; 44%, 22% with 400 μL), adjusting the composition formulas of citronella oil had a synergistic effect, for more effective repellent against Ae. aegypti.

Mosquitoes transmit a variety of pathogens that have devastating consequences for global public and veterinary health. Despite their capacity to serve as vectors, these insects have a robust capacity to respond to invading organisms with strong cellular and humoral immune responses. In Egypt, Aedes caspius (Pallas, 1771) has been suspected to act as a bridge vector of Rift Valley Fever virus between animals and humans. Microscopic analysis of Ae. caspius hemolymph revealed the presence of phagocytic cells called granulocytes. We further evaluated cellular immune responses produced by Ae. caspius as a result of exposure to a Gram-negative, and Gram-positive bacterium, and to latex beads. After challenge, a rapid and strong phagocytic response against either a natural or synthetic invader was evident. Hemocyte integrity in bacteria-inoculated mosquitoes was not morphologically affected. The number of circulating granulocytes decreased with age, reducing the overall phagocytic capacity of mosquitoes over time. The magnitude and speed of the phagocytic response suggested that granulocytes act as an important force in the battle against foreign invaders, as has been characterized in other important mosquito vector species.

A mosquito strain of Aerdes albopictus,HAmAalG0,from Huntsville,Alabama,USA,showed a normal susceptibility and low tolerance to permethrin and resmethrin (pyrethroid insecticides) compared to a susceptible Ikaken strain,even though these pyrethroid insecticides have been used in the field for a long period of time in Alabama.Recently,we treated HAmAalG0 in the laboratory with permethrin for five generations and detected no significant change in the level of resistance to permethrin in the selected mosquitoes,HAmAalG5,compared with the parental strain HAmAalG0. We then examined the allelic expression at the L-to-F kdr site of the sodium channel gene in the Aedesmosquitoes to address our hypothesis that the L-to-F kdr mutation was not present in HAmAalG0 and HAmAalG5 mosquitoes. We found that every tested individual in Ikaken,HAmAalG0,and HAmAalG5 populations expressed a codon of CTA at the L-to-F kdr site encoding Leu,strongly corresponding to their susceptibility to insecticides.

Nectar feeding by males and females of the mosquitoAedes provocans was studied at a site near Belleville, Ontario, Canada. Canada plum, Prunus nigra, and especially pin cherry, P. pensylvanica, bloomed contemporaneously with the emergence of Ae. provocans and were important nectar sources for adult mosquitoes during their first week of life. Blossoms of P. pensylvanica shielded for 24 h from foragers produced an average of 0.14 mg of sugar (approximately 2.3J). This nectar was avidly sought by both sexes of Ae. provocans; > 97% of the blossoms were visited by mosquitoes in the first few days of blooming. Young adult mosquitoes were found on blossoms at all hours of the day and night; feeding on P. nigra was strongly eocrepuscular, whereas on P. pensylvanica feeding was much less strongly periodic. Adults foraged for nectar in an energy-conserving, pedestrian strategy, devoting 56% (females) and 68% (males) of their time on blossoms to nectar feeding during foraging bouts that lasted a median of 5.3 min. Both sexes sought nectar soon after emergence--males before they had completed hypopygial rotation or swarmed, and females before mating or host seeking. Female Ae. provocans sought nectar in all stages of oogenesis, but primarily at the initiation of a gonotrophic cycle. Energy stores in the crop averaged 18J per female, with a distribution that depended on gonotrophic age and parity.

La Crosse virus (LACV), a leading cause of arboviral encephalitis in children in the United States, is emerging in Appalachia. For local arboviral surveillance, mosquitoes were tested. LACV RNA was detected and isolated from Aedes japonicus mosquitoes. These invasive mosquitoes may significantly affect LACV range expansion and dynamics. PMID:25811131

Zika virus (ZIKV) remained obscure until the recent explosive outbreaks in French Polynesia (2013-2014) and South America (2015-2016). Phylogenetic studies have shown that ZIKV has evolved into African and Asian lineages. The Asian lineage of ZIKV was responsible for the recent epidemics in the Americas. However, the underlying mechanisms through which ZIKV rapidly and explosively spread from Asia to the Americas are unclear. Non-structural protein 1 (NS1) facilitates flavivirus acquisition by mosquitoes from an infected mammalian host and subsequently enhances viral prevalence in mosquitoes. Here we show that NS1 antigenaemia determines ZIKV infectivity in its mosquito vector Aedes aegypti, which acquires ZIKV via a blood meal. Clinical isolates from the most recent outbreak in the Americas were much more infectious in mosquitoes than the FSS13025 strain, which was isolated in Cambodia in 2010. Further analyses showed that these epidemic strains have higher NS1 antigenaemia than the FSS13025 strain because of an alanine-to-valine amino acid substitution at residue 188 in NS1. ZIKV infectivity was enhanced by this amino acid substitution in the ZIKV FSS13025 strain in mosquitoes that acquired ZIKV from a viraemic C57BL/6 mouse deficient in type I and II interferon (IFN) receptors (AG6 mouse). Our results reveal that ZIKV evolved to acquire a spontaneous mutation in its NS1 protein, resulting in increased NS1 antigenaemia. Enhancement of NS1 antigenaemia in infected hosts promotes ZIKV infectivity and prevalence in mosquitoes, which could have facilitated transmission during recent ZIKV epidemics.

México has cities (e.g., México City and Puebla City) located at elevations > 2,000 m and above the elevation ceiling below which local climates allow the dengue virus mosquito vector Aedes aegypti to proliferate. Climate warming could raise this ceiling and place high-elevation cities at risk for dengue virus transmission. To assess the elevation ceiling for Ae. aegypti and determine the potential for using weather/climate parameters to predict mosquito abundance, we surveyed 12 communities along an elevation/climate gradient from Veracruz City (sea level) to Puebla City (∼2,100 m). Ae. aegypti was commonly encountered up to 1,700 m and present but rare from 1,700 to 2,130 m. This finding extends the known elevation range in México by > 300 m. Mosquito abundance was correlated with weather parameters, including temperature indices. Potential larval development sites were abundant in Puebla City and other high-elevation communities, suggesting that Ae. aegypti could proliferate should the climate become warmer. PMID:22987656

Full Text Available A detailed analysis of Prangos pabularia Lindl. (Apiaceae fruit oil was performed by gas chromatography (GC-FID and gas chromatography-mass spectrometry (GC-MS. Bicyclogermacrene (21%, (Z- b -ocimene (19%, a -humulene (8%, a -pinene (8% and spathulenol (6% were the main constituents of the oil. One compound with 1.8% at RI 3420 remained unidentified or tentatively identified as suberosin from the Wiley GC-MS Library. T he assumed compound, suberosin was synthesized in two steps and its structure was confirmed by 1D NMR and GC- MS analyses. As part of our continued research to discover new chemicals for use in mosquito control agents as repellents and larvicides, suberosin and its parent compound coumarin were investigated for the mosquito biting deterrent and larvicidal activity against Aedes aegypti. Both suberosin and coumarin showed biting deterrent activity but the activity was lower than the positive control, DEET (N,N-diethyl-3-methylbenzamide. In larval bioassays, suberosin with LC 50 value of 8.1 ppm was significantly more toxic than c oumarin (LC 50 = 49.6 ppm at 24-h post treatment. These results indicate that suberosin may be useful for use as mosquito larvicidal agent .

In tropical and sub-tropical regions, the mosquitoAedes aegypti is the major vector for the virus causing dengue, a serious public health issue in these areas. Through ongoing NSF- and NASA-funded studies, field surveys of Aedes aegypti and an integrated modeling approach are being used to improve our understanding of the potential range of the mosquito to expand toward heavily populated high elevation areas such as Mexico City under various climate change and socio-economic scenarios. This work serves three primary objectives: (1) Employ NASA remotely-sensed data to supplement the environmental monitoring and modeling component of the project. These data -- for example, surface temperature, precipitation, vegetation indices, soil moisture and elevation -- are critical for understanding the habitat necessary for mosquito survival and abundance; (2) Implement training sessions to instruct scientists and students from Mexico and the U.S. on how to use remote sensing and implement the NASA SERVIR Regional Visualization and Monitoring System; (3) Employ the SERVIR framework to optimize the dissemination of key project results in order to increase their societal relevance and benefits in developing climate adaptation strategies. Field surveys of larval, pupal and adult Aedes aegypti, as well as detailed physical and social household characteristics, were conducted in the summers of 2011and 2012 at geographic scales from the household to the community along a transect from sea level to 2400 m ASL. These data are being used in models to estimate Aedes aegypti habitat suitability. In 2011, Aedes aegypti were identified at an elevation of over 2150 m in Puebla, the highest elevation at which this species has been observed.

In tropical and sub ]tropical regions, the mosquitoAedes aegypti is the major vector for the virus causing dengue, a serious public health issue in these areas. Through ongoing NSF- and NASA-funded studies, field surveys of Aedes aegypti and an integrated modeling approach are being used to improve our understanding of the potential range of the mosquito to expand toward heavily populated high elevation areas such as Mexico City under various climate change and socio ]economic scenarios. This work serves three primary objectives: (1) Employ NASA remotely-sensed data to supplement the environmental monitoring and modeling component of the project. These data-- for example, surface temperature, precipitation, vegetation indices, soil moisture and elevation-- are critical for understanding the habitat necessary for mosquito survival and abundance; (2) Implement training sessions to instruct scientists and students from Mexico and the U.S. on how to use remote sensing and implement the NASA SERVIR Regional Visualization and Monitoring System; (3) Employ the SERVIR framework to optimize the dissemination of key project results in order to increase their societal relevance and benefits in developing climate adaptation strategies. Field surveys of larval, pupal and adult Aedes aegypti, as well as detailed physical and social household characteristics, were conducted in the summers of 2011and 2012 at geographic scales from the household to the community along a transect from sea level to 2400 m ASL. These data are being used in models to estimate Aedes aegypti habitat suitability. In 2011, Aedes aegypti were identified at an elevation of over 2150 m in Puebla, the highest elevation at which this species has been observed.

The Asian tiger mosquito, Aedes albopictus, is a highly invasive mosquito species that transmits chikungunya and dengue. This species overwinters as diapausing eggs in temperate climates. Early diapause termination may be a beneficial strategy for winter mosquito control; however, a mechanism to terminate the diapause process using chemicals is not known. We tested the hypothesis that a hormonal imbalance caused by the administration of juvenile hormone analog would terminate egg diapause in A. albopictus. We tested the insect growth regulator pyriproxyfen on all developmental stages to identify a susceptible stage for diapause termination. We found that pyriproxyfen treatment of mosquito eggs terminated embryonic diapause. The highest rates of diapause termination were recorded in newly deposited (78.9%) and fully embryonated (74.7%) eggs at 0.1 and 1 ppm, respectively. Hatching was completed earlier in newly deposited eggs (25-30 days) compared to fully embryonated eggs (71-80 days). The combined mortality from premature diapause termination and ovicidal activity was 98.2% in newly deposited and >98.9% in fully embryonated eggs at 1 ppm. The control diapause eggs did not hatch under diapausing conditions. Pyriproxyfen exposure to larvae, pupae and adults did not prevent the females from ovipositing diapausing eggs. There was no effect of pyriproxyfen on diapausing egg embryonic developmental time. We also observed mortality in diapausing eggs laid by females exposed to pyriproxyfen immediately after blood feeding. There was no mortality in eggs laid by females that survived larval and pupal exposures. In conclusion, diapausing eggs were the more susceptible to pyriproxyfen diapause termination compared to other life stages. This is the first report of diapause termination in A. albopictus with a juvenile hormone analog. We believe our findings will be useful in developing a new control strategy against overwintering mosquito populations.

Full Text Available The Asian tiger mosquito, Aedes albopictus, is a highly invasive mosquito species that transmits chikungunya and dengue. This species overwinters as diapausing eggs in temperate climates. Early diapause termination may be a beneficial strategy for winter mosquito control; however, a mechanism to terminate the diapause process using chemicals is not known. We tested the hypothesis that a hormonal imbalance caused by the administration of juvenile hormone analog would terminate egg diapause in A. albopictus. We tested the insect growth regulator pyriproxyfen on all developmental stages to identify a susceptible stage for diapause termination. We found that pyriproxyfen treatment of mosquito eggs terminated embryonic diapause. The highest rates of diapause termination were recorded in newly deposited (78.9% and fully embryonated (74.7% eggs at 0.1 and 1 ppm, respectively. Hatching was completed earlier in newly deposited eggs (25-30 days compared to fully embryonated eggs (71-80 days. The combined mortality from premature diapause termination and ovicidal activity was 98.2% in newly deposited and >98.9% in fully embryonated eggs at 1 ppm. The control diapause eggs did not hatch under diapausing conditions. Pyriproxyfen exposure to larvae, pupae and adults did not prevent the females from ovipositing diapausing eggs. There was no effect of pyriproxyfen on diapausing egg embryonic developmental time. We also observed mortality in diapausing eggs laid by females exposed to pyriproxyfen immediately after blood feeding. There was no mortality in eggs laid by females that survived larval and pupal exposures. In conclusion, diapausing eggs were the more susceptible to pyriproxyfen diapause termination compared to other life stages. This is the first report of diapause termination in A. albopictus with a juvenile hormone analog. We believe our findings will be useful in developing a new control strategy against overwintering mosquito populations.

The Asian tiger mosquito, Aedes albopictus, is a highly invasive mosquito species that transmits chikungunya and dengue. This species overwinters as diapausing eggs in temperate climates. Early diapause termination may be a beneficial strategy for winter mosquito control; however, a mechanism to terminate the diapause process using chemicals is not known. We tested the hypothesis that a hormonal imbalance caused by the administration of juvenile hormone analog would terminate egg diapause in A. albopictus. We tested the insect growth regulator pyriproxyfen on all developmental stages to identify a susceptible stage for diapause termination. We found that pyriproxyfen treatment of mosquito eggs terminated embryonic diapause. The highest rates of diapause termination were recorded in newly deposited (78.9%) and fully embryonated (74.7%) eggs at 0.1 and 1 ppm, respectively. Hatching was completed earlier in newly deposited eggs (25–30 days) compared to fully embryonated eggs (71–80 days). The combined mortality from premature diapause termination and ovicidal activity was 98.2% in newly deposited and >98.9% in fully embryonated eggs at 1 ppm. The control diapause eggs did not hatch under diapausing conditions. Pyriproxyfen exposure to larvae, pupae and adults did not prevent the females from ovipositing diapausing eggs. There was no effect of pyriproxyfen on diapausing egg embryonic developmental time. We also observed mortality in diapausing eggs laid by females exposed to pyriproxyfen immediately after blood feeding. There was no mortality in eggs laid by females that survived larval and pupal exposures. In conclusion, diapausing eggs were the more susceptible to pyriproxyfen diapause termination compared to other life stages. This is the first report of diapause termination in A. albopictus with a juvenile hormone analog. We believe our findings will be useful in developing a new control strategy against overwintering mosquito populations. PMID:26090954

Mosquito-borne diseases have an economic impact, including loss in commercial and labor outputs, particularly in countries with tropical and subtropical climates; however, no part of the world is free from vector-borne diseases. In mosquito control programs, botanical origin may have the potential to be used successfully as eggs, larvae, and adult. The larvicidal, ovicidal, and repellent activities of crude benzene and ethyl acetate extracts of leaf of Ervatamia coronaria and Caesalpinia pulcherrima were assayed for their toxicity against three important vector mosquitoes, viz., Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus (Diptera: Culicidae). The larval mortality was observed after 24 h of exposure. All extracts showed moderate larvicidal effects; however, the highest larval mortality was found in benzene extract of E. coronaria against the larvae of Anopheles Stephensi, Aedes aegypti, and Culex quinquefasciatus with the LC(50) and LC(90) values were 79.08, 89.59, and 96.15 ppm and 150.47, 166.04, and 174.10 ppm, respectively. Mean percent hatchability of the ovicidal activity was observed 48 h posttreatment. The percent hatchability was inversely proportional to the concentration of extract and directly proportional to the eggs. The leaf extract of E. coronaria was found to be most effective than Caesalpinia pulcherrima against eggs/egg rafts of three vector mosquitoes. For E. coronaria, the benzene extract exerted 300, 250, and 200 ppm against Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus, respectively. The results of the repellent activity of benzene and ethyl acetate extract of E. coronaria and Caesalpinia pulcherrima plants at three different concentrations of 1.0, 2.5, and 5.0 mg/cm(2) were applied on skin of fore arm in man and exposed against adult female mosquitoes. In this observation, these two plant crude extracts gave protection against mosquito bites without any allergic reaction to the test person, and also, the

Aedes-borne viruses are responsible for high-impact neglected tropical diseases and unpredictable outbreaks such as the ongoing Zika epidemics. Aedesmosquitoes spread different arboviruses such as Dengue virus (DENV), Chikungunya virus (CHIKV), and Zika virus, among others, and are responsible for the continuous emergence and reemergence of these pathogens. These viruses have complex transmission cycles that include two hosts, namely the Aedesmosquito as a vector and susceptible vertebrate hosts. Human infection with arboviruses causes diseases that range from subclinical or mild to febrile diseases, encephalitis, and hemorrhagic fever. Infected mosquitoes do not show detectable signs of disease, even though the virus maintains a lifelong persistent infection. The infection of the Aedesmosquito by viruses involves a molecular crosstalk between cell and viral proteins. An understanding of how mosquito vectors and viruses interact is of fundamental interest, and it also offers novel perspectives for disease control. In recent years, mass spectrometry (MS)-based strategies in combination with bioinformatics have been successfully applied to identify and quantify global changes in cellular proteins, lipids, peptides, and metabolites in response to viral infection. Although the information about proteomics in the Aedesmosquito is limited, the information that has been reported can set up the basis for future studies. This review reflects how MS-based approaches have extended our understanding of Aedesmosquito biology and the development of DENV and CHIKV infection in the vector. Finally, this review discusses future challenges in the field.

The effect of spraying a mixture of the insect growth regulator (IGR) pyriproxyfen (1 mg/liter) and either 1% boric acid sugar bait or eugenol sugar bait on croton petra plants (Codiaeum variegatum L.) was evaluated against the container-inhabiting mosquito, Aedes albopictus (Skuse). Treatments were applied to plants and evaluated against adult and larval Ae. albopictus in the laboratory through contact and wash off experiments, respectively. The control treatment lacked an active ingredient and were treated with an attractive sugar bait. The plants treated with attractive toxic sugar baits plus the IGR resulted in 60-100% mortality of laboratory-reared adult Ae. albopictus. The pyriproxyfen solutions collected from the plant wash experiment resulted in 80-100% emergence inhibition to the exposed third- and fourth-instar larvae, compared with the untreated control. Attractive toxic sugar baits mixed with the IGR not only provide effective control of adult mosquitoes, but also provide additional control of larvalmosquitoes after being washed off from the treated plants.

The quality of breeding sites is reflected through the pupal productivity and the life history traits of Aedesmosquitoes. Using nutrient reserves and pupal productivity of Aedes as indicators, the larval habitats including sewage drains were characterized to highlight the habitat expansion and vector management. The pupae and adults collected from the containers and sewage drains were characterized in terms of biomass and nutrient reserves and the data were subjected to three way factorial ANOVA. Discriminant function analyses were performed to highlight the differences among the habitats for sustenance of Aedesmosquitoes. Survey of larval habitats from the study area revealed significant differences (PAedes among the habitats and months. Despite sewage drains being comparatively less utilized for breeding, the pupae were of higher biomass with corresponding adults having longer wings in contrast to other habitats. The nutrient reserve of the adults emerging from pupae of sewage drains was significantly higher (Paegypti and Ae. albopictus, sewage drains were equally congenial habitat as were plastic, porcelain and earthen habitats. Availability of Aedes immature in sewage drains poses increased risk of dengue, and thus vector control programme should consider inclusion of sewage drains as breeding habitat of dengue vector mosquitoes.

Full Text Available Abstract Background Bacillus thuringiensis var. israelensis (Bti is a natural larvalmosquito pathogen producing pore-forming toxins targeting the midgut of Diptera larvae. It is used worldwide for mosquito control. Resistance mechanisms of an Aedes aegypti laboratory strain selected for 30 generations with field-collected leaf litter containing Bti toxins were investigated in larval midguts at two levels: 1. gene transcription using DNA microarray and RT-qPCR and 2. differential expression of brush border membrane proteins using DIGE (Differential In Gel Electrophoresis. Results Several Bti Cry toxin receptors including alkaline phosphatases and N-aminopeptidases and toxin-binding V-ATPases exhibited altered expression levels in the resistant strain. The under-expression of putative Bti-receptors is consistent with Bt-resistance mechanisms previously described in Lepidoptera. Four soluble metalloproteinases were found under-transcribed together with a drastic decrease of metalloproteinases activity in the resistant strain, suggesting a role in resistance by decreasing the amount of activated Cry toxins in the larval midgut. Conclusions By combining transcriptomic and proteomic approaches, we detected expression changes at nearly each step of the ingestion-to-infection process, providing a short list of genes and proteins potentially involved in Bti-resistance whose implication needs to be validated. Collectively, these results open the way to further functional analyses to better characterize Bti-resistance mechanisms in mosquitoes.

Full Text Available The mosquito immune system is involved in pathogen-elicited defense responses. The NF-κB factors REL1 and REL2 are downstream transcription activators of Toll and IMD immune pathways, respectively. We have used genome-wide microarray analyses to characterize fat-body-specific gene transcript repertoires activated by either REL1 or REL2 in two transgenic strains of the mosquitoAedes aegypti. Vitellogenin gene promoter was used in each transgenic strain to ectopically express either REL1 (REL1+ or REL2 (REL2+ in a sex, tissue, and stage specific manner. There was a significant change in the transcript abundance of 297 (79 up- and 218 down-regulated and 299 (123 up- and 176 down-regulated genes in fat bodies of REL1+ and REL2+, respectively. Over half of the induced genes had predicted functions in immunity, and a large group of these was co-regulated by REL1 and REL2. By generating a hybrid transgenic strain, which ectopically expresses both REL1 and REL2, we have shown a synergistic action of these NF-κB factors in activating immune genes. The REL1+ immune transcriptome showed a significant overlap with that of cactus (RNAi-depleted mosquitoes (50%. In contrast, the REL2+ -regulated transcriptome differed from the relatively small group of gene transcripts regulated by RNAi depletion of a putative inhibitor of the IMD pathway, caspar (35 up- and 140 down-regulated, suggesting that caspar contributes to regulation of a subset of IMD-pathway controlled genes. Infections of the wild type Ae. aegypti with Plasmodium gallinaceum elicited the transcription of a distinct subset of immune genes (76 up- and 25 down-regulated relative to that observed in REL1+ and REL2+ mosquitoes. Considerable overlap was observed between the fat body transcriptome of Plasmodium-infected mosquitoes and that of mosquitoes with transiently depleted PIAS, an inhibitor of the JAK-STAT pathway. PIAS gene silencing reduced Plasmodium proliferation in Ae. aegypti, indicating

Full Text Available Abstract Background The evolutionary importance of horizontal gene transfer (HGT from Wolbachia endosymbiotic bacteria to their eukaryotic hosts is a topic of considerable interest and debate. Recent transfers of genome fragments from Wolbachia into insect chromosomes have been reported, but it has been argued that these fragments may be on an evolutionary trajectory to degradation and loss. Results We have discovered a case of HGT, involving two adjacent genes, between the genomes of Wolbachia and the currently Wolbachia-uninfected mosquitoAedes aegypti, an important human disease vector. The lower level of sequence identity between Wolbachia and insect, the transcription of all the genes involved, and the fact that we have identified homologs of the two genes in another Aedes species (Ae. mascarensis, suggest that these genes are being expressed after an extended evolutionary period since horizontal transfer, and therefore that the transfer has functional significance. The association of these genes with Wolbachia prophage regions also provides a mechanism for the transfer. Conclusion The data support the argument that HGT between Wolbachia endosymbiotic bacteria and their hosts has produced evolutionary innovation.

Functional Characterization of the Octenol Receptor Neuron on the Maxillary Palps of the Yellow Fever Mosquito, Aedes aegypti Alan J. Grant, Joseph C...Dickens JC (2011) Functional Characterization of the Octenol Receptor Neuron on the Maxillary Palps of the Yellow Fever Mosquito, Aedes aegypti . PLoS...palps. Both sexes of mosquitoes possess basiconic sensilla that contain three neurons; in Aedes aegypti these sensilla number about 35 in females and 21

In a laboratory study, the insect growth regulator, cyromazine, exerted a high level of biological activity on Aedes aegypti and Culex quinquefasciatus treated in the 4th larval instar. At 1.5 and 1.0 ppm this IGR produced 97 and 99% inhibition of emergence in adult Ae. aegypti, respectively. In Cx. quinquefasciatus, there was 99% inhibition at 1 ppm and complete inhibition at 1.5 ppm. The overall pupal mortality was higher than larval or adult stages of both species. This material induced different types of morphogenetic abnormalities in pupae and adults of the 2 species similar to those induced by other IGRs. However, most abnormalities were observed in the pupal stage. Adverse effects were not detected on the 4 mosquito predator species during the acute or posttreatment tests.

Urban parks are areas designated for human recreation but also serve as shelter and refuge for populations of several species of native fauna, both migratory and introduced. In Brazil, the effect of annual climate variations on Aedes aegypti and dengue epidemics in large cities like São Paulo is well known, but little is known about how such variations can affect the diversity of mosquito vectors in urban parks and the risk of disease transmission by these vectors. This study investigates the influence of larval habitats and seasonal factors on the diversity and abundance of Culicidae fauna in Anhanguera Park, one of the largest remaining green areas in the city of São Paulo. Species composition and richness and larval habitats were identified. Seasonality (cold-dry and hot-rainy periods) and year were considered as explanatory variables and the models selection approach was developed to investigate the relationship of these variables with mosquito diversity and abundance. A total of 11,036 specimens from 57 taxa distributed in 13 genera were collected. Culex nigripalpus, Cx. quinquefasciatus and Aedes albopictus were the most abundant species. Bamboo internodes and artificial breeding sites showed higher abundance, while ponds and puddles showed greater richness. Significant relationships were observed between abundance and seasonality, with a notable increase in the mosquitos abundance in the warm-rainy periods. The Shannon and Berger-Parker indices were related with interaction between seasonality and year, however separately these predictors showed no relationship with ones. The increased abundance of mosquitoes in warm-rainy months and the fact that some of the species are epidemiologically important increase not only the risk of pathogen transmission to people who frequent urban parks but also the nuisance represented by insect bites. The findings of this study highlight the importance of knowledge of culicid ecology in green areas in urban environments.

The female Aedes aegypti mosquito is an important vector for several tropical and subtropical diseases such as dengue, chikungunya, and Zika and yellow fever. The disease viruses infect the mosquito and subsequently spread to the salivary glands after which the viruses can be transmitted to humans with probing or feeding by the mosquito. Omics systems sciences offer the opportunity to characterize vectors and can inform disease surveillance, vector control and development of innovative diagnostics, personalized medicines, vaccines, and insecticide targets. Using high-resolution mass spectrometry, we performed an analysis of the A. aegypti salivary gland proteome. The A. aegypti proteome resulted in acquisition of 83,836 spectra. Upon searches against the protein database of the A. aegypti, these spectra were assigned to 5417 unique peptides, belonging to 1208 proteins. To the best of our knowledge, this is the largest set of proteins identified in the A. aegypti salivary gland. Of note, 29 proteins were involved in immunity-related pathways in salivary glands. A subset of these proteins is known to interact with disease viruses. Another 15 proteins with signal cleavage site were found to be secretory in nature, and thus possibly playing critical roles in blood meal ingestion. These findings provide a baseline to advance our understanding of vector-borne diseases and vector-pathogen interactions before virus transmission in global health, and might therefore enable future design and development of virus-blocking strategies and novel molecular targets in the mosquito vector A. aegypti.

Four antennae-specific proteins (AaegOBP1, AaegOBP2, AaegOBP3, and AaegASP1) were isolated from the yellow fever mosquito, Aedes aegypti and their full-length cDNAs were cloned. RT-PCR indicated that they are expressed in female and, to a lesser extent, in male antennae, but not in control tissues (legs). AaegOBP1 and AaegOBP3 showed significant similarity to previously identified mosquito odorant-binding proteins (OBPs) in cysteine spacing pattern and sequence. Two of the isolated proteins have a total of eight cysteine residues. The similarity of the spacing pattern of the cysteine residues and amino acid sequence to those of previously identified olfactory proteins suggests that one of the cysteine-rich proteins (AaegOBP2) is an OBP. The other (AaegASP1) did not belong to any group of known OBPs. Structural analyses indicate that six of the cysteine residues in AaegOBP2 are linked in a similar pattern to the previously known cysteine pairing in OBPs, i.e., Cys-24 Cys-55, Cys-51 Cys-104, Cys-95 Cys-113. The additional disulfide bridge, Cys-38 Cys-125, knits the extended C-terminal segment of the protein to a predicted α2-helix. As indicated by circular dichroism (CD) spectra, the extra rigidity seems to prevent the predicted formation of a C-terminal α-helix at low pH.

Pyriproxyfen, a juvenile hormone analog insect growth regulator (IGR), is a recommended insecticide for the control of container-inhabiting mosquitoes. The effects of eight container substrates (three plastics, wood, tire rubber, clay pot, concrete, and glass) were studied on reduction of bio-efficacy of pyriproxyfen for Aedes albopictus (Skuse) in the laboratory. Insect growth regulator-bioassay of third instars showed significant increase of LC50 with some substrates compared with control probably because of pyriproxyfen adsorption to the substrate. The LC50 for the control substrate (glass mug) was 0.029 microg/liter. In comparison to the control, LC50 increased 150.1 times with tire rubber (4.354 microg/liter) and 8-10 times with wood and high-density polyethylene plastic containers. Slight increases of LC50 were detected for clay pots, polypropylene, and polystyrene plastics (1.9-2.7 times). Pyriproxyfen efficacy with concrete substrate was higher than with glass containers, which might be caused by high pH. The pH of water with wood (5.6) and concrete (9.8) substrates differed from the other substrates tested (6.3-7.1). There was no pupal mortality at pH 5-11 without pyriproxyfen; however, interaction of pH (5-11) with pyriproxyfen (0.05 microg/liter) showed additive effects at both low and high pH values. Different substrates have variable impacts on pyriproxyfen efficacy, which might be associated with the failure of larval control strategies for the container mosquitoes under certain field conditions.

This study describes a laboratory investigation on the use of the insect-pathogenic fungus Metarhizium anisopliae against adult Aedes aegypti and Ae. albopictus mosquitoes. At a dosage of 1.6 × 1010 conidia/m2, applied on material that served as a mosquito resting site, an average of 87.1 ± 2.65% of

This study describes a laboratory investigation on the use of the insect-pathogenic fungus Metarhizium anisopliae against adult Aedes aegypti and Ae. albopictus mosquitoes. At a dosage of 1.6 × 1010 conidia/m2, applied on material that served as a mosquito resting site, an average of 87.1 ± 2.65% of

Since a safe and effective mass vaccination program against dengue fever is not presently available, a good way to prevent and control dengue outbreaks depends mainly on controlling the mosquito vectors. Aedes aegypti mosquito populations can be monitored and reduced by using ovitraps baited with organic infusions. A series of laboratory experiments were conducted which demonstrated that the bacteria in bamboo leaf infusion produce volatile attractants and contact chemical stimulants attractive to the female mosquitoes. The results showed that the female mosquitoes laid most of their eggs (59.9 ± 8.1 vs 2.9 ± 2.8 eggs, PAedes mosquitoes. Aedes aegypti laid significantly more eggs (63.3 ± 6.5 vs 3.1 ± 2.4 eggs, P<0.001) in bacteria suspension compared to sterile R2A medium. Our results suggest microbial activity has a role in the production of odorants that mediate the oviposition response of gravid mosquitoes.

Aedes aegypti larvae in both areas under vector control and no vector control. METHODS: World Health Organization standard bioassays for diagnostic concentration and multiple concentrations were performed in mosquito larvae collected in an area under no vector control (Campinas, SP and an area under vector chemical control (Campo Grande, MS, in Brazil. RESULTS: Potential resistance to a diagnostic concentration of temephos (DC=0.04 ppm was registered for an Ae. aegypti larval population collected in Campinas. Multiple concentration tests confirmed the larvae resistance, with 24.5% of them surviving at the 0.0125 ppm concentration. Bioassays with the organophosphate fenitrothion (DC=0.08 ppm and pyrethroid cypermethrin (DC=0.01 ppm in the same population revealed their susceptibility to these agents. Bioassays carried out in an Ae. aegypti larval population collected in Campo Grande showed their susceptibility to temephos (DC=0.04 ppm and cypermethrin (DC=0.01 ppm. LC50 and LC95 for cypermethrin (CE25, cyfluthrin (CE5, betacyfluthrin (SC1.25 and propoxur (CE20 were determined for Ae. aegypti . Using the Rockefeller standard strain values, ratios of resistance were estimated: 2.9, 2.2, 2.4 and 1.3 for LC50 and 3.5, 2.6, 3.9 and 1.3 for LC95, respectively. CONCLUSION: The findings reinforce the need for routinely monitoring pesticide efficacy as a very important step in vector control management programs.

OR I GI N AL C ONTR I BUTI O N Gene silencing in adult Aedes aegypti mosquitoes through oral delivery of double-stranded RNA M. R. Coy1, N. D...we tested whether such an approach could be used in the yellow fever mosquito, Aedes aegypti . Using a non-specific dsRNA construct, we found that...adult Ae. aegypti ingested dsRNA through this method and that the ingested dsRNA can be recovered from the mosquitoes post-feeding. Through the feeding of

Copepods from La Plata, Argentina were investigated to characterize the local community of larvivorous copepods inhabiting mosquito breeding sites and to identify new predator species of the mosquitoes which occur in artificial containers, Aedes aegypti and Culex pipiens. Diversity of larvivorous cyclopoids was highest in permanent pools. Predation by sex and age, selectivity on mosquito species, and daily predation rate during five days were studied for Acanthocyclops robustus, Diacyclops uruguayensis, Macrocyclops albidus and Mesocyclops longisetus. Female copepods presented the highest predatory capacity. No predatory preference for mosquito species was found. According to overall predation potential, copepods were ranked as follows: D. uruguayensis biological control agents for mosquitoes.

Full Text Available Chikungunya virus (CHIKV is an emerging arbovirus transmitted to humans by mosquitoes such as Aedes albopictus. To be transmitted, CHIKV must replicate in the mosquito midgut, then disseminate in the hemocele and infect the salivary glands before being released in saliva. We have developed a standardized protocol to visualize viral particles in the mosquito salivary glands using transmission electron microscopy. Here we provide direct evidence for CHIKV replication and storage in Ae. albopictus salivary glands.

Chikungunya virus (CHIKV) is an emerging arbovirus transmitted to humans by mosquitoes such as Aedes albopictus. To be transmitted, CHIKV must replicate in the mosquito midgut, then disseminate in the hemocele and infect the salivary glands before being released in saliva. We have developed a standardized protocol to visualize viral particles in the mosquito salivary glands using transmission electron microscopy. Here we provide direct evidence for CHIKV replication and storage in Ae. albopictus salivary glands.

Mosquito attractants and spatial repellents hold great promise in controlling mosquito pests. In assessing the effectiveness of mosquito attractants and repellents, a good olfactometer system, and optimized testing conditions, are essential. In this research, we demonstrated the usefulness of an olfactometer system, and optimized testing conditions for Aedes albopictus (Diptera: Culicidae). We found no significant difference in the biting activity of the insect between 8:00 and 22:00. Further...

The mating efficiencies of field-collected and laboratory-colonized Aedes triseriatus (Say) (Diptera: Culicidae) female mosquitoes transovarially-infected or uninfected with La Crosse virus (LACV) were compared. The females were placed in cages with age-matched males, and the insemination rates were determined daily by detection of sperm in the spermathecae. LACV-infected mosquitoes typically mated more quickly than uninfected mosquitoes. LACV load was not correlated with increased insemination. PMID:19769048

Copepods from La Plata, Argentina were investigated to characterize the local community of larvivorous copepods inhabiting mosquito breeding sites and to identify new predator species of the mosquitoes which occur in artificial containers, Aedes aegypti and Culex pipiens. Diversity of larvivorous cyclopoids was highest in permanent pools. Predation by sex and age, selectivity on mosquito species, and daily predation rate during five days were studied for Acanthocyclops robustus, Diacyclops ur...

Copepods from La Plata, Argentina were investigated to characterize the local community of larvivorous copepods inhabiting mosquito breeding sites and to identify new predator species of the mosquitoes which occur in artificial containers, Aedes aegypti and Culex pipiens. Diversity of larvivorous cyclopoids was highest in permanent pools. Predation by sex and age, selectivity on mosquito species, and daily predation rate during five days were studied for Acanthocyclops robustus, Diacyclops ur...

The Colombo City in Sri Lanka is experiencing tremendous development and construction of multiple storey buildings and high rise apartments. The change in housing types and microhabitats might have altered the flight and breeding behaviour of Aedesmosquito population. This study was carried out to determine the vertical dispersal and abundance of Aedesmosquitoes in multiple storey buildings in the Colombo district, with respect to abiotic factors such as rainfall, humidity and wind speed. Hence, this study is of paramount importance, particularly for planning and implementation of control measures against Aedesmosquitoes. An ovitrap based study was carried out at four selected multiple storey buildings in four residential areas located in Colombo, Sri Lanka, from August to December 2013. Results were analyzed using four indices; ovitrap index, mean number of larvae, mean number of eggs and mean number of larvae per ovipaddle. The results implied that Aedesmosquitoes could be found in different elevations from ground floor to the highest floor (130 ft). There was a significant difference between height and ovitrap index (povitrap index, mean number of larvae and mean number of larvae per ovipaddle) also displayed higher values, i.e. 13.19 ± 2.98%, 1.366 ± 0.527, and 2.070 ± 0.421%, respectively. Abiotic factors such as wind speed, coastal nature, etc. displayed a significant effect to the vertical dispersal of Aedesmosquitoes (p0.05) with height which indicates high abundance of Aedesmosquitoes at higher floors. Abiotic factors also seemed to cause significant effect to the vertical dispersal of Aedesmosquitoes in high rise buildings.

Co-infections may modify parasite transmission opportunities directly as a consequence of interactions in the within-host environment, but also indirectly through changes in host life history. Furthermore, host and parasite traits are sensitive to the abiotic environment with variable consequences for parasite transmission in co-infections. We investigate how co-infection of the mosquitoAedes aegypti with two microsporidian parasites (Vavraia culicis and Edhazardia aedis) at two levels of larval food availability affects parasite transmission directly, and indirectly through effects on host traits. In a laboratory infection experiment, we compared how co-infection, at low and high larval food availability, affected the probability of infection, within-host growth and the transmission potential of each parasite, compared to single infections. Horizontal transmission was deemed possible for both parasites when infected hosts died harbouring horizontally transmitting spores. Vertical transmission was judged possible for E. aedis when infected females emerged as adults. We also compared the total input number of spores used to seed infections with output number, in single and co-infections for each parasite. The effects of co-infection on parasite fitness were complex, especially for V. culicis. In low larval food conditions, co-infection increased the chances of mosquitoes dying as larvae or pupae, thus increasing opportunities for V. culicis' horizontal transmission. However, co-infection reduced larval longevity and hence time available for V. culicis spore production. Overall, there was a negative net effect of co-infection on V. culicis, whereby the number of spores produced was less than the number used to seed infection. Co-infections also negatively affected horizontal transmission of the more virulent parasite, E. aedis, through reduced longevity of pre-adult hosts. However, its potential transmission suffered less relative to V. culicis. Our results

Full Text Available Intervention measures to control the transmission of vector-borne diseases include control of the vector population. In mosquito control, synthetic insecticides used against both the larvae (larvicides and adults (adulticides create numerous problems, such as environmental pollution, insecticide resistance and toxic hazards to humans. In the present study, a bacterial pesticide, Bacillus sphaericus (Bs G3-IV, was used to control the dengue and filarial vectors, Aedes aegypti and Culex quinquefasciatus. Bacillus sphaericus (Bs G3-IV was very effective against Aedes aegypti and Culex quinquefasciatus, showing significant larval mortality. Evaluated lethal concentrations (LC50 and LC90 were age-dependent, with early instars requiring a lower concentration compared with later stages of mosquitoes. Culex quinquefasciatus was more susceptible to Bacillus sphaericus (Bs G3-IV than was Aedes aegypti. Fecundity rate was highly reduced after treatment with different concentrations of Bacillus sphaericus (Bs G3-IV. Larval and pupal longevity both decreased after treatment with Bacillus sphaericus (Bs G3-IV, total number of days was lower in the B. sphaericus treatments compared with the control. Our results show the bacterial pesticide Bacillus sphaericus (Bs G3-IV to be an effective mosquito control agent that can be used for more integrated pest management programs.

To assess the larvicidal and repellent potential of the essential oil extracted from the leaves of peppermint plant, Mentha piperita (M. piperita) against the larval and adult stages of Aedes aegypti (Ae. Aegypti). The larvicidal potential of peppermint oil was evaluated against early fourth instar larvae of Ae. aegypti using WHO protocol. The mortality counts were made after 24 and 48 h, and LC50 and LC90 values were calculated. The efficacy of peppermint oil as mosquito repellent was assessed using the human-bait technique. The measured area of one arm of a human volunteer was applied with the oil and the other arm was applied with ethanol. The mosquito bites on both the arms were recorded for 3 min after every 15 min. The experiment continued for 3 h and the percent protection was calculated. The essential oil extracted from M. piperita possessed excellent larvicidal efficiency against dengue vector. The bioassays showed an LC50 and LC90 value of 111.9 and 295.18 ppm, respectively after 24 h of exposure. The toxicity of the oil increased 11.8% when the larvae were exposed to the oil for 48 h. The remarkable repellent properties of M. piperita essential oil were established against adults Ae. aegypti. The application of oil resulted in 100% protection till 150 min. After next 30 min, only 1-2 bites were recorded as compared with 8-9 bites on the control arm. The peppermint essential oil is proved to be efficient larvicide and repellent against dengue vector. Further studies are needed to identify the possible role of oil as adulticide, oviposition deterrent and ovicidal agent. The isolation of active ingredient from the oil could help in formulating strategies for mosquito control.

Objective To assess the larvicidal and repellent potential of the essential oil extracted from the leaves of peppermint plant, Mentha piperita (M. piperita) against the larval and adult stages of Aedes aegypti (Ae. Aegypti). Methods The larvicidal potential of peppermint oil was evaluated against early fourth instar larvae of Ae. aegypti using WHO protocol. The mortality counts were made after 24 and 48 h, and LC50 and LC90 values were calculated. The efficacy of peppermint oil as mosquito repellent was assessed using the human-bait technique. The measured area of one arm of a human volunteer was applied with the oil and the other arm was applied with ethanol. The mosquito bites on both the arms were recorded for 3 min after every 15 min. The experiment continued for 3 h and the percent protection was calculated. Results The essential oil extracted from M. piperita possessed excellent larvicidal efficiency against dengue vector. The bioassays showed an LC50 and LC90 value of 111.9 and 295.18 ppm, respectively after 24 h of exposure. The toxicity of the oil increased 11.8% when the larvae were exposed to the oil for 48 h. The remarkable repellent properties of M. piperita essential oil were established against adults Ae. aegypti. The application of oil resulted in 100% protection till 150 min. After next 30 min, only 1-2 bites were recorded as compared with 8-9 bites on the control arm. Conclusions The peppermint essential oil is proved to be efficient larvicide and repellent against dengue vector. Further studies are needed to identify the possible role of oil as adulticide, oviposition deterrent and ovicidal agent. The isolation of active ingredient from the oil could help in formulating strategies for mosquito control. PMID:23569733

Objective:To assess the larvicidal and repellent potential of the essential oil extracted from the leaves of peppermint plant,Mentha piperita (M. piperita) against the larval and adult stages of Aedes aegypti (Ae. Aegypti).Methods: The larvicidal potential of peppermint oil was evaluated against early fourth instar larvae ofAe. aegypti usingWHO protocol. The mortality counts were made after 24 and48 h, and LC50 and LC90values were calculated. The efficacy of peppermint oil as mosquito repellent was assessed using the human-bait technique. The measured area of one arm of a human volunteer was applied with the oil and the other arm was applied with ethanol. The mosquito bites on both the arms were recorded for3 min after every15 min. The experiment continued for 3 h and the percent protection was calculated.Results:The essential oil extracted fromM. piperita possessed excellent larvicidal efficiency against dengue vector. The bioassays showed an LC50 and LC90 value of111.9 and295.18 ppm, respectively after24 h of exposure. The toxicity of the oil increased11.8% when the larvae were exposed to the oil for48 h. The remarkable repellent properties ofM. piperita essential oil were established against adults Ae. aegypti. The application of oil resulted in100% protection till150 min. After next30min, only1-2 bites were recorded as compared with8-9 bites on the control arm.Conclusions:The peppermint essential oil is proved to be efficient larvicide and repellent against dengue vector. Further studies are needed to identify the possible role of oil as adulticide, oviposition deterrent and ovicidal agent. The isolation of active ingredient from the oil could help in formulating strategies for mosquito control.

In China, the prevention and control of Zika virus disease has been a public health threat since the first imported case was reported in February 2016. To determine the vector competence of potential vector mosquito species, we experimentally infected Aedes aegypti, Ae. albopictus, and Culex quinquefasciatus mosquitoes and determined infection rates, dissemination rates, and transmission rates. We found the highest vector competence for the imported Zika virus in Ae. aegypti mosquitoes, some susceptibility of Ae. albopictus mosquitoes, but no transmission ability for Cx. quinquefasciatus mosquitoes. Considering that, in China, Ae. albopictus mosquitoes are widely distributed but Ae. aegypti mosquito distribution is limited, Ae. albopictus mosquitoes are a potential primary vector for Zika virus and should be targeted in vector control strategies. PMID:28430562

Two insular settlements (Kampung Pulau Ketam and Kampung Sungai Lima) were selected to study the population dynamics of Aedes aegypti and Aedes albopictus mosquitoes, vectors of dengue and chikungunya infections. Ovitrap surveillance was conducted between October 2007 and October 2008. There was an inverse negative association between ovitrap index and rainfall at the time of collection, probably because rainfall increased the number of available oviposition sites. Rainfall and ovitrap index were positively associates the 25th day after rainfall occurred. A minor, second peak was observed from the 38th to the 42nd day. The first peak was consistent with the minimum 18-day period between the hatching of eggs to the first oviposition. The second minor peak could be due to the second gonotrophic cycle of the female mosquitoes. Rainfall is an important environmental factor associated with Aedes breeding at the study sites.

Full Text Available BACKGROUND: Wolbachia infections confer protection for their insect hosts against a range of pathogens including bacteria, viruses, nematodes and the malaria parasite. A single mechanism that might explain this broad-based pathogen protection is immune priming, in which the presence of the symbiont upregulates the basal immune response, preparing the insect to defend against subsequent pathogen infection. A study that compared natural Wolbachia infections in Drosophila melanogaster with the mosquito vector Aedes aegypti artificially transinfected with the same strains has suggested that innate immune priming may only occur in recent host-Wolbachia associations. This same study also revealed that while immune priming may play a role in viral protection it cannot explain the entirety of the effect. METHODOLOGY/FINDINGS: Here we assess whether the level of innate immune priming induced by different Wolbachia strains in A. aegypti is correlated with the degree of protection conferred against bacterial pathogens. We show that Wolbachia strains wMel and wMelPop, currently being tested for field release for dengue biocontrol, differ in their protective abilities. The wMelPop strain provides stronger, more broad-based protection than wMel, and this is likely explained by both the higher induction of immune gene expression and the strain-specific activation of particular genes. We also show that Wolbachia densities themselves decline during pathogen infection, likely as a result of the immune induction. CONCLUSIONS/SIGNIFICANCE: This work shows a correlation between innate immune priming and bacterial protection phenotypes. The ability of the Toll pathway, melanisation and antimicrobial peptides to enhance viral protection or to provide the basis of malaria protection should be further explored in the context of this two-strain comparison. This work raises the questions of whether Wolbachia may improve the ability of wild mosquitoes to survive pathogen

Aedes albopictus and Ae. japonicus are two of the most widespread invasive mosquito species that have recently become established in western Europe. Both species are associated with the transmission of a number of serious diseases and are projected to continue their spread in Europe. In the present study, we modelled the habitat suitability for both species under current and future climatic conditions by means of an Ensemble forecasting approach. We additionally compared the modelled MAXENT niches of Ae. albopictus and Ae. japonicus regarding temperature and precipitation requirements. Both species were modelled to find suitable habitat conditions in distinct areas within Europe: Ae. albopictus within the Mediterranean regions in southern Europe, Ae. japonicus within the more temperate regions of central Europe. Only in few regions, suitable habitat conditions were projected to overlap for both species. Whereas Ae. albopictus is projected to be generally promoted by climate change in Europe, the area modelled to be climatically suitable for Ae. japonicus is projected to decrease under climate change. This projection of range reduction under climate change relies on the assumption that Ae. japonicus is not able to adapt to warmer climatic conditions. The modelled MAXENT temperature niches of Ae. japonicus were found to be narrower with an optimum at lower temperatures compared to the niches of Ae. albopictus. Species distribution models identifying areas with high habitat suitability can help improving monitoring programmes for invasive species currently in place. However, as mosquito species are known to be able to adapt to new environmental conditions within the invasion range quickly, niche evolution of invasive mosquito species should be closely followed upon in future studies.

Dengue fever is among the most widespread vector-borne infectious diseases. The primary vector of dengue is the Aedes aegypti mosquito. Ae. aegypti is prevalent in the tropics and sub-tropics and is closely associated with human habitats outside its native range of Africa. While long established in the southeastern United States of America where dengue is re-emerging, breeding populations have never been reported from California until the summer of 2013. Using 12 highly variable microsatellite loci and a database of reference populations, we have determined that the likely source of the California introduction is the southeastern United States, ruling out introductions from abroad, from the geographically closer Arizona or northern Mexico populations, or an accidental release from a research laboratory. The power to identify the origin of new introductions of invasive vectors of human disease relies heavily on the availability of a panel of reference populations. Our work demonstrates the importance of generating extensive reference databases of genetically fingerprinted human-disease vector populations to aid public health efforts to prevent the introduction and spread of vector-borne diseases.

Objective: To investigate the prevalence of container breeding mosquitoes with emphasis on the seasonality and larval habitats of Aedes aegypti (Ae. aegypti) in Makkah City, adjoining an environmental monitoring and dengue incidence. Methods: Monthly visits were performed between April 2008 and March 2009 to randomly selected houses. During each visit, mosquito larvae were collected from indoors and outdoors containers by either dipping or pipetting. Mosquitoes were morphologically identified. Data on temperature, relative humidity, rain/precipitations during the survey period was retrieved from governmental sources and analyzed. Results: The city was warmer in dry season (DS) than wet season (WS). No rain occurred at all during DS and even precipitations did fall, wetting events were much greater during WS. Larval survey revealed the co-breeding of Aedes, Culex and Anopheles in a variety of artificial containers in and around homes. 32109 larvae representing 1st , 2nd, 3rd, and 4th stages were collected from 22618 container habitats. Culicines was far the commonest and Aedes genus was as numerous as the Culex population. Ae. aegypti larval abundance exhibited marked temporal variations, overall, being usually more abundant during WS. Ten types of artificial containers were found with developing larvae. 70% of these habitats were located indoors. 71.42% of indoor containers were permanent and 28.58% was semi-permanent during WS. Cement tanks was the only container type permanent during DS. Ae. aegypti larval indices (CI, HI, BI) recorded were greater during WS. Conclusions: Taken together, these results indicate a high risk of dengue transmission in the holy city.

Full Text Available Abstract Background In western Kenya, malaria remains one of the major health problems and its control remains an important public health measure. Malaria control is by either use of drugs to treat patients infected with malaria parasites or by controlling the vectors. Vector control may target the free living adult or aquatic (larval stages of mosquito. The most commonly applied control strategies target indoor resting mosquitoes. However, because mosquitoes spend a considerable time in water, targeting the aquatic stages can complement well with existing adult control measures. Methods Larval source management (LSM of malaria vectors was examined in two villages i.e. Fort Ternan and Lunyerere, with the aim of testing strategies that can easily be accessed by the affected communities. Intervention strategies applied include environmental management through source reduction (drainage of canals, land levelling or by filling ditches with soil, habitat manipulation (by provision of shading from arrow root plant, application of Bacillus thuringiensis var israelensis (Bti and the use of predatory fish, Gambusia affinis. The abundance of immature stages of Anopheles and Culex within intervention habitats was compared to that within non-intervention habitats. Results The findings show that in Fort Ternan no significant differences were observed in the abundance of Anopheles early and late instars between intervention and non-intervention habitats. In Lunyerere, the abundance of Anopheles early instars was fifty five times more likely to be present within non-intervention habitats than in habitats under drainage. No differences in early instars abundance were observed between non-intervention and habitats applied with Bti. However, late instars had 89 % and 91 % chance of being sampled from non-intervention rather than habitats under drainage and those applied with Bti respectively. Conclusion Most of these interventions were applied in habitats

Interventions targeting adult mosquitoes are used to combat transmission of vector-borne diseases, including dengue. Without available vaccines, targeting the primary vector, Aedes aegypti, is essential to prevent transmission. Older mosquitoes (≥ 7 days) are of greatest epidemiological significance due to the 7-day extrinsic incubation period of the virus. Age-grading of female mosquitoes is necessary to identify post-intervention changes in mosquito population age structure. We developed models using near-infrared spectroscopy (NIRS) to age-grade adult female Ae. aegypti. To determine if diet affects the ability of NIRS models to predict age, two identical larval groups were fed either fish food or infant cereal. Adult females were separated and fed sugar water ± blood, resulting in four experimental groups. Females were killed 1, 4, 7, 10, 13, or 16 days postemergence. The head/thorax of each mosquito was scanned using a near-infrared spectrometer. Scans from each group were analyzed, and multiple models were developed using partial least squares regression. The best model included all experimental groups, and positively predicted the age group (< or ≥ 7 days) of 90.2% mosquitoes. These results suggest both larval and adult diets can affect the ability of NIRS models to accurately assign age categories to female Ae. aegypti.

Reverse genetics is a powerful tool for understanding gene functions and their interactions in the mosquito innate immunity. We took the transgenic approach, in combination with the RNA interference (RNAi) technique, to elucidate the role of mosquito REL1, a homolog of Drosophila Dorsal, in regulation of Toll immune pathway in the mosquitoAedes aegypti. By transforming the mosquitoes with ΔREL1-A or a double-stranded RNA construct of REL1 driven by the female fat body-specific vitellogenin (...

This ovitrap study examined the effects of altitude above sea level, tree species, and tree trunk diameter on the distribution of eggs of the 2 tree hole mosquitoes, Aedes hendersoni and Ae. triseriatus. Only tree species and trunk diameter affected the distribution significantly. Aedes hendersoni eggs were found more frequently associated with trees of border and sunny habitat, while Ae. triseriatus eggs were more frequently found in association with trees of mesic habitat. Oviposition of Ae. hendersoni occurred more often at trees with smaller diameter at breast height than did Ae. triseriatus. These differences in ovipositing frequency appear to be related to the microhabitat associated with different sample sites.

The mosquitoAedes aegypti is the principal vector of dengue and yellow fever flaviviruses. Temephos is an organophosphate insecticide used globally to suppress Ae. aegypti larval populations but resistance has evolved in many locations. Quantitative Trait Loci (QTL) controlling temephos survival in Ae. aegypti larvae were mapped in a pair of F3 advanced intercross lines arising from temephos resistant parents from Solidaridad, México and temephos susceptible parents from Iquitos, Peru. Two sets of 200 F3 larvae were exposed to a discriminating dose of temephos and then dead larvae were collected and preserved for DNA isolation every two hours up to 16 hours. Larvae surviving longer than 16 hours were considered resistant. For QTL mapping, single nucleotide polymorphisms (SNPs) were identified at 23 single copy genes and 26 microsatellite loci of known physical positions in the Ae. aegypti genome. In both reciprocal crosses, Multiple Interval Mapping identified eleven QTL associated with time until death. In the Solidaridad×Iquitos (SLD×Iq) cross twelve were associated with survival but in the reciprocal IqxSLD cross, only six QTL were survival associated. Polymorphisms at acetylcholine esterase (AchE) loci 1 and 2 were not associated with either resistance phenotype suggesting that target site insensitivity is not an organophosphate resistance mechanism in this region of México. Temephos resistance is under the control of many metabolic genes of small effect and dispersed throughout the Ae. aegypti genome.

Reverse genetics is a powerful tool for understanding gene functions and their interactions in the mosquito innate immunity. We took the transgenic approach, in combination with the RNA interference (RNAi) technique, to elucidate the role of mosquito REL1, a homolog of Drosophila Dorsal, in regulation of Toll immune pathway in the mosquitoAedes aegypti. By transforming the mosquitoes with DeltaREL1-A or a double-stranded RNA construct of REL1 driven by the female fat body-specific vitellogenin (Vg) promoter with the pBac[3xP3-EGFP, afm] vector, we generated two different transgenic mosquito strains, one with overexpressed AaREL1 and the second with AaREL1 knockdown. Both strains had a single copy of the respective transgene, and the expression in both transgenic mosquitoes was highly activated by blood feeding. Vg-DeltaREL1-A transgenic mosquitoes activate Toll immune pathway in the fat body by blood feeding. The overexpression of both isoforms, AaREL1-A and AaREL1-B, in Vg-DeltaREL1-A transgenic mosquitoes resulted in the concomitant activation of Aedes Spätzle1A and Serpin-27A, independent of septic injury. The same phenotype was observed in the mosquitoes with RNAi knockdown of an Aedes homolog to Drosophila cactus, an IkappaB inhibitor of Drosophila Toll pathway. The effect of the transgenic RNAi knockdown of AaREL1 on mosquito innate immunity was revealed by increased susceptibility to the entomopathogenic fungus Beauveria bassiana and the reduced induction of Spz1A and Serpin-27A gene expression after fungal challenge. These results have proven that AaREL1 is a key downstream regulator of Toll immune pathway in the mosquito A. aegypti.

The presented paper deal with the mosquito control in the Padova district (Italy). Control strategies and operative planning are reported, together with an example of census schedule. Further development in aedes albopictus control are pointed out.

Interpretation & conclusion: The study concludes that there was high MIR in the identified localities of low and medium income groups. Estimation of MIR in a female Aedesmosquito in the existing arsenals for dengue surveillance would be an added advantage for early warning of dengue outbreak. The presence of infected mosquitoes in identified localities of Delhi was alarming and require rigorous vector surveillance so that the severe outbreaks can be prevented.

In the Western Hemisphere, Zika virus is thought to be transmitted primarily by Aedes aegypti mosquitoes. To determine the extent to which Ae. albopictus mosquitoes from the United States are capable of transmitting Zika virus and the influence of virus dose, virus strain, and mosquito species on vector competence, we evaluated multiple doses of representative Zika virus strains in Ae. aegypti and Ae. albopictus mosquitoes. Virus preparation (fresh vs. frozen) significantly affected virus infectivity in mosquitoes. We calculated 50% infectious doses to be 6.1–7.5 log10 PFU/mL; minimum infective dose was 4.2 log10 PFU/mL. Ae. albopictus mosquitoes were more susceptible to infection than Ae. aegypti mosquitoes, but transmission efficiency was higher for Ae. aegypti mosquitoes, indicating a transmission barrier in Ae. albopictus mosquitoes. Results suggest that, although Zika virus transmission is relatively inefficient overall and dependent on virus strain and mosquito species, Ae. albopictus mosquitoes could become major vectors in the Americas. PMID:28430564

The chimeric yellow fever (YF) 17D-dengue type 2 (ChimeriVax-DEN2) vaccine virus developed by Acambis, Inc. (Cambridge, MA) contains the prM and E genes of wild-type (wt) dengue 2 (DEN-2) (strain PUO-218) virus in the YF vaccine virus (strain 17D) backbone. The potential of ChimeriVax-DEN2 virus to infect and be transmitted by Aedes aegypti, the principal DEN and YF virus mosquito vector, and Aedes albopictus, a species that occurs in areas of active transmission of YF and DEN viruses, was evaluated. Mosquitoes were intrathoracically (IT) inoculated with virus or were fed a virus-laden blood meal, and the replication kinetics of ChimeriVax-DEN2 were compared with the wt DEN-2 and YF 17D vaccine viruses. Replication of YF 17D virus is attenuated in cultured Ae. albopictus C6/36 mosquito cells and in Ae. aegypti and Ae. albopictus mosquitoes. Growth of ChimeriVax-DEN2 virus similarly was restricted in C6/36 cells and in mosquitoes. ChimeriVax-DEN2 replicated in 56% of IT inoculated Ae. aegypti, and virus disseminated to head tissue in 36%, with a mean viral titer of 1.8 log10 PFU/mosquito. Of mosquitoes, 16% of Ae. aegypti and 24% of Ae. albopictus were infected 14 days after a blood meal containing ChimeriVax-DEN2, but virus did not disseminate to head tissue. In contrast, DEN-2 replicated in all IT inoculated and orally infected Ae. aegypti (mean titer 5.5 log10 PFU/mosquito), and virus disseminated to head tissue in 95%. Of Ae. albopictus, 84% were infected after a blood meal containing DEN-2 virus; dissemination occurred in 36%. Replication of ChimeriVax-DEN2 virus in mosquitoes corresponded to that of YF 17D vaccine virus, which is restricted in its ability to infect and replicate in mosquitoes. Therefore, transmission of ChimeriVax-DEN2 virus by vector mosquitoes is unlikely.

The aim of the present study was to determine the vertical distribution and abundance of Aedesmosquitoes in multiple storey buildings in Selangor and Kuala Lumpur, Malaysia. Ovitrap surveillance was conducted for 4 continuous weeks in multiple storey buildings in 4 residential areas located in Selangor [Kg. Baiduri (KB)] and Kuala Lumpur [Student Hostel of University of Malaya (UM), Kg. Kerinchi (KK) and Hang Tuah (HT)]. The results implied that Aedesmosquitoes could be found from ground floor to highest floor of multiple storey buildings and data from different elevation did not show significant difference. Ovitrap index for UM, KB, HT and KK ranged from 0 - 29.17%, 0 - 55.56%, 8.33 - 83.33% and 0 - 91.17% respectively. Aedes aegypti and Aedes albopictus were found breeding in HT, KK and KB; while only Ae. albopictus was obtained from UM. The results indicate that the invasion of Aedesmosquitoes in high-rise apartments could facilitate the transmission of dengue virus and new approaches to vector control in this type of residential area should be developed.

Chikungunya virus (CHIKV) was isolated from 12 febrile humans in Yucatan, Mexico, in 2015. One patient was co-infected with dengue virus type 1. Two additional CHIKV isolates were obtained from Aedes aegypti mosquitoes collected in the homes of patients. Phylogenetic analysis showed that the CHIKV isolates belong to the Asian lineage.

Chikungunya virus (CHIKV) was isolated from 12 febrile humans in Yucatan, Mexico, in 2015. One patient was co-infected with dengue virus type 1. Two additional CHIKV isolates were obtained from Aedes aegypti mosquitoes collected in the homes of patients. Phylogenetic analysis showed that the CHIKV isolates belong to the Asian lineage. PMID:27347760

Aedes aegypti (L.) (Diptera: Culicidae) transmit pathogens to humans, leading to diseases such as yellow fever and dengue fever. Repellents and insecticides are two common interventions to reduce mosquito biting and thereby disease risk. However, overreliance on a chemical or class of chemicals c...

In 1958, the Pan American Health Organization declared that Brazil had successfully eradicated the mosquitoAedes aegypti, responsible for the transmission of yellow fever, dengue fever, chikungunya, and Zika virus. Yet in 2016 the Brazilian minister of health described the situation of dengue fever as "catastrophic." Discussing the recent epidemic of Zika virus, which amplified the crisis produced by the persistence of dengue fever, Brazil's president declared in January 2016 that "we are in the process of losing the war against the mosquitoAedes aegypti." I discuss the reasons for the failure to contain Aedes in Brazil and the consequences of this failure. A longue durée perspective favors a view of the Zika epidemic that does not present it as a health crisis to be contained with a technical solution alone but as a pathology that has the persistence of deeply entrenched structural problems and vulnerabilities.

Full Text Available Lentic freshwater systems including those inhabited by aquatic stages of mosquitoes derive most of their carbon inputs from terrestrial organic matter mainly leaf litter. The leaf litter is colonized by microbial communities that provide the resource base for mosquito larvae. While the microbial biomass associated with different leaf species in container aquatic habitats is well documented, the taxonomic composition of these microbes and their response to common environmental stressors is poorly understood. We used indoor aquatic microcosms to determine the abundances of major taxonomic groups of bacteria in leaf litters from seven plant species and their responses to low concentrations of four pesticides with different modes of action on the target organisms; permethrin, malathion, atrazine and glyphosate. We tested the hypotheses that leaf species support different quantities of major taxonomic groups of bacteria and that exposure to pesticides at environmentally relevant concentrations alters bacterial abundance and community structure in mosquitolarval habitats. We found support for both hypotheses suggesting that leaf litter identity and chemical contamination may alter the quality and quantity of mosquito food base (microbial communities in larval habitats. The effect of pesticides on microbial communities varied significantly among leaf types, suggesting that the impact of pesticides on natural microbial communities may be highly complex and difficult to predict. Collectively, these findings demonstrate the potential for detritus composition within mosquitolarval habitats and exposure to pesticides to influence the quality of mosquitolarval habitats.

Background: Considering the disadvantages of chemical insecticides, we aimed to evaluate Vorticella parasites for control of mosquito larvae of Anopheles stephensi and Aedes aegypti at different larval stages.Methods: Vorticella sp infected mosquito larvae were crushed in the 0.85% saline and homogenized well to get Vorti­cella in suspension. The effects of Vorticella sp infections on larval development were investigated by inoculat­ing protozoan on different larval instars of An. stephensi a...

Background: Considering the disadvantages of chemical insecticides, we aimed to evaluate Vorticella parasites for control of mosquito larvae of Anopheles stephensi and Aedes aegypti at different larval stages. Methods: Vorticella sp infected mosquito larvae were crushed in the 0.85% saline and homogenized well to get Vorticella in suspension. The effects of Vorticella sp infections on larval development were investigated by inoculating protozoan on different larval instars of An. stephensi an...

Biotic interactions involving mosquito larvae are context dependent, with effects of interactions on populations altered by ecological conditions. Relative impacts of competition and predation change across a gradient of habitat size and permanence. Asymmetrical competition is common and ecological context changes competitive advantage, potentially facilitating landscape-level coexistence of competitors. Predator effects on mosquito populations sometimes depend on habitat structure and on emergent effects of multiple predators, particularly interference among predators. Nonlethal effects of predators on mosquito oviposition, foraging, and life history are common, and their consequences for populations and for mosquito-borne disease are poorly understood. Context-dependent beneficial effects of detritus shredders on mosquitoes occur in container habitats, but these interactions appear to involve more than simple resource modification by shredders. Investigations of context-dependent interactions among mosquito larvae will yield greater understanding of mosquito population dynamics and provide useful model systems for testing theories of context dependence in communities.

Several diseases are associated to the mosquito-human interaction. Mosquitoes are the carriers of severe and well-known illnesses such as malaria, arboviral encephalitis, dengue fever, chikungunya fever, West Nile virus and yellow fever. These diseases produce significant morbidity and mortality in humans and livestock around the world. The present investigation was undertaken to study the ovicidal, larvicidal and adulticidal activities of crude hexane, ethyl acetate, benzene, chloroform and methanol extracts of root of Asparagus racemosus were assayed for their toxicity against three important vector mosquitoes, viz., Culex quinquefasciatus, Aedes aegypti and Anopheles stephensi (Diptera: Culicidae). The mean percent hatchability of the eggs was observed after 48 h post-treatment. The percent hatchability was inversely proportional to the concentration of extract and directly proportional to the eggs. All the five solvent extracts showed moderate ovicidal activity; however, the methanol extract showed the highest ovicidal activity. The methanol extract of Asparagus racemosus against Culex quinquefasciatus, Aedes aegypti and Anopheles stephensi exerted 100% mortality (zero hatchability) at 375, 300 and 225 ppm, respectively. Control eggs showed 99-100% hatchability. The larval mortality was observed after 24 h of exposure. All extracts showed moderate larvicidal effects; however, the highest larval mortality was found in methanol extract of root of Asparagus racemosus against the larvae of Culex quinquefasciatus, Aedes aegypti and Anopheles stephensi with the LC50 and LC90 values were 115.13, 97.71 and 90.97 ppm and 210.96, 179.92, and 168.82 ppm, respectively. The adult mortality was observed after 24 h recovery period. The plant crude extracts showed dose-dependent mortality. At higher concentrations, the adult showed restless movement for some times with abnormal wagging and then died. Among the extracts tested, the highest adulticidal activity was observed in

Zika virus, already widely distributed in Africa and Asia, was recently reported in two Northeastern Brazilian: State of Bahia and State of Rio Grande do Norte, and one Southeastern: State of São Paulo. This finding adds a potentially noxious virus to a list of several other viruses that are widely transmitted by Aedes (Stegomyia) aegypti and Aedes (Stegomyia) albopictus in Brazil. The pathology and epidemiology, including the distribution and vectors associated with Zika virus, are reviewed. This review is focused on viruses transmitted by Aedes (Stegomyia) mosquitoes, including dengue, Chikungunya, Zika, Mayaro, and yellow fever virus, to emphasize the risks of occurrence for these arboviruses in Brazil and neighboring countries. Other species of Aedes (Stegomyia) are discussed, emphasizing their involvement in arbovirus transmission and the possibility of adaptation to environments modified by human activities and introduction in Brazil.

Full Text Available Abstract Zika virus, already widely distributed in Africa and Asia, was recently reported in two Northeastern Brazilian: State of Bahia and State of Rio Grande do Norte, and one Southeastern: State of São Paulo. This finding adds a potentially noxious virus to a list of several other viruses that are widely transmitted by Aedes (Stegomyia aegypti and Aedes (Stegomyia albopictus in Brazil. The pathology and epidemiology, including the distribution and vectors associated with Zika virus, are reviewed. This review is focused on viruses transmitted by Aedes (Stegomyia mosquitoes, including dengue, Chikungunya, Zika, Mayaro, and yellow fever virus, to emphasize the risks of occurrence for these arboviruses in Brazil and neighboring countries. Other species of Aedes (Stegomyia are discussed, emphasizing their involvement in arbovirus transmission and the possibility of adaptation to environments modified by human activities and introduction in Brazil.

The existing literature about HCV association with, and replication in mosquitoes is extremely poor. To fill this gap, we performed cellular investigations aimed at exploring (i) the capacity of HCV E1E2 glycoproteins to bind on Aedesmosquito cells and (ii) the ability of HCV serum particles (HCVsp) to replicate in these cell lines. First, we used purified E1E2 expressing baculovirus-derived HCV pseudo particles (bacHCVpp) so we could investigate their association with mosquito cell lines from Aedes aegypti (Aag-2) and Aedes albopictus (C6/36). We initiated a series of infections of both mosquito cells (Ae aegypti and Ae albopictus) with the HCVsp (Lat strain - genotype 3) and we observed the evolution dynamics of viral populations within cells over the course of infection via next-generation sequencing (NGS) experiments. Our binding assays revealed bacHCVpp an association with the mosquito cells, at comparable levels obtained with human hepatocytes (HepaRG cells) used as a control. In our infection experiments, the HCV RNA (+) were detectable by RT-PCR in the cells between 21 and 28 days post-infection (p.i.). In human hepatocytes HepaRG and Ae aegypti insect cells, NGS experiments revealed an increase of global viral diversity with a selection for a quasi-species, suggesting a structuration of the population with elimination of deleterious mutations. The evolutionary pattern in Ae albopictus insect cells is different (stability of viral diversity and polymorphism). These results demonstrate for the first time that natural HCV could really replicate within Aedesmosquitoes, a discovery which may have major consequences for public health as well as in vaccine development.

Background In the past ten years, the Indian Ocean region has been the theatre of severe epidemics of chikungunya and dengue. These outbreaks coincided with a high increase in populations of Aedes albopictus that outcompete its sister taxon Aedes aegypti in most islands sampled. The objective of this work was to update the entomological survey of the two Aedes species in the island of Madagascar which has to face these arboviroses. Methods The sampling of Aedesmosquitoes was conducted during two years, from October 2007 to October 2009, in fifteen localities from eight regions of contrasting climates. Captured adults were identified immediately whereas immature stages were bred until adult stage for determination. Phylogenetic analysis was performed using two mtDNA genes, COI and ND5 and trees were constructed by the maximum likelihood (ML) method with the gene time reversible (GTR) model. Experimental infections with the chikungunya virus strain 06.21 at a titer of 107.5 pfu/mL were performed to evaluate the vector competence of field-collected mosquitoes. Disseminated infection rates were measured fourteen days after infection by immunofluorescence assay performed on head squashes. Results The species Aedes aegypti was detected in only six sites in native forests and natural reserves. In contrast, the species Aedes albopictus was found in 13 out of the 15 sites sampled. Breeding sites were mostly found in man-made environments such as discarded containers, used tires, abandoned buckets, coconuts, and bamboo cuts. Linear regression models showed that the abundance of Ae. albopictus was significantly influenced by the sampling region (F = 62.00, p Madagascar, but the markers used were not discriminant enough to discern Malagasy populations. The experimental oral infection method showed that six Ae. albopictus populations exhibited high dissemination infection rates for chikungunya virus ranging from 98 to 100%. Conclusion In Madagascar, Ae. albopictus has extended

Full Text Available Abstract Background In the past ten years, the Indian Ocean region has been the theatre of severe epidemics of chikungunya and dengue. These outbreaks coincided with a high increase in populations of Aedes albopictus that outcompete its sister taxon Aedes aegypti in most islands sampled. The objective of this work was to update the entomological survey of the two Aedes species in the island of Madagascar which has to face these arboviroses. Methods The sampling of Aedesmosquitoes was conducted during two years, from October 2007 to October 2009, in fifteen localities from eight regions of contrasting climates. Captured adults were identified immediately whereas immature stages were bred until adult stage for determination. Phylogenetic analysis was performed using two mtDNA genes, COI and ND5 and trees were constructed by the maximum likelihood (ML method with the gene time reversible (GTR model. Experimental infections with the chikungunya virus strain 06.21 at a titer of 107.5 pfu/mL were performed to evaluate the vector competence of field-collected mosquitoes. Disseminated infection rates were measured fourteen days after infection by immunofluorescence assay performed on head squashes. Results The species Aedes aegypti was detected in only six sites in native forests and natural reserves. In contrast, the species Aedes albopictus was found in 13 out of the 15 sites sampled. Breeding sites were mostly found in man-made environments such as discarded containers, used tires, abandoned buckets, coconuts, and bamboo cuts. Linear regression models showed that the abundance of Ae. albopictus was significantly influenced by the sampling region (F = 62.00, p -16 and period (F = 36.22, p = 2.548 × 10-13, that are associated with ecological and climate variations. Phylogenetic analysis of the invasive Ae. albopictus distinguished haplotypes from South Asia and South America from those of Madagascar, but the markers used were not discriminant enough

A longitudinal study was conducted in Manaus, Brazil, to monitor changes of adult Aedes aegypti (L.) abundance. The objectives were to compare mosquito collections of two trap types, to characterise temporal changes of the mosquito population, to investigate the influence of meteorological variables on mosquito collections and to analyse the association between mosquito collections and dengue incidence. Mosquito monitoring was performed fortnightly using MosquiTRAPs (MQT) and BG-Sentinel (BGS) traps between December 2008-June 2010. The two traps revealed opposing temporal infestation patterns, with highest mosquito collections of MQTs during the dry season and highest collections of BGS during the rainy seasons. Several meteorological variables were significant predictors of mosquito collections in the BGS. The best predictor was the relative humidity, lagged two weeks (in a positive relationship). For MQT, only the number of rainy days in the previous week was significant (in a negative relationship). The correlation between monthly dengue incidence and mosquito abundance in BGS and MQT was moderately positive and negative, respectively. Catches of BGS traps reflected better the dynamic of dengue incidence. The findings help to understand the effects of meteorological variables on mosquito infestation indices of two different traps for adult dengue vectors in Manaus. PMID:25494470

The survival characteristics of the mosquitoAedes aegypti affect transmission rates of dengue because transmission requires infected mosquitoes to survive long enough for the virus to infect the salivary glands. Mosquito survival is assumed to be high in tropical, dengue endemic, countries like Vietnam. However, the survival rates of wild populations of mosquitoes are seldom measured due the difficulty of predicting mosquito age. Hon Mieu Island in central Vietnam is the site of a pilot release of Ae. aegypti infected with a strain of Wolbachia pipientis bacteria (wMelPop) that induces virus interference and mosquito life-shortening. We used the most accurate mosquito age grading approach, transcriptional profiling, to establish the survival patterns of the mosquito population from the population age structure. Furthermore, estimations were validated on mosquitoes released into a large semi-field environment consisting of an enclosed house, garden and yard to incorporate natural environmental variability. Mosquito survival was highest during the dry/cool (January-April) and dry/hot (May-August) seasons, when 92 and 64% of Hon Mieu mosquitoes had survived to an age that they were able to transmit dengue (12 d), respectively. This was reduced to 29% during the wet/cool season from September to December. The presence of Ae. aegypti older than 12 d during each season is likely to facilitate the observed continuity of dengue transmission in the region. We provide season specific Ae. aegypti survival models for improved dengue epidemiology and evaluation of mosquito control strategies that aim to reduce mosquito survival to break the dengue transmission cycle.

Full Text Available The survival characteristics of the mosquitoAedes aegypti affect transmission rates of dengue because transmission requires infected mosquitoes to survive long enough for the virus to infect the salivary glands. Mosquito survival is assumed to be high in tropical, dengue endemic, countries like Vietnam. However, the survival rates of wild populations of mosquitoes are seldom measured due the difficulty of predicting mosquito age. Hon Mieu Island in central Vietnam is the site of a pilot release of Ae. aegypti infected with a strain of Wolbachia pipientis bacteria (wMelPop that induces virus interference and mosquito life-shortening. We used the most accurate mosquito age grading approach, transcriptional profiling, to establish the survival patterns of the mosquito population from the population age structure. Furthermore, estimations were validated on mosquitoes released into a large semi-field environment consisting of an enclosed house, garden and yard to incorporate natural environmental variability. Mosquito survival was highest during the dry/cool (January-April and dry/hot (May-August seasons, when 92 and 64% of Hon Mieu mosquitoes had survived to an age that they were able to transmit dengue (12 d, respectively. This was reduced to 29% during the wet/cool season from September to December. The presence of Ae. aegypti older than 12 d during each season is likely to facilitate the observed continuity of dengue transmission in the region. We provide season specific Ae. aegypti survival models for improved dengue epidemiology and evaluation of mosquito control strategies that aim to reduce mosquito survival to break the dengue transmission cycle.

Sexual size dimorphism (SSD) in insects often accompanies a sexual difference in development time, sexual bimaturism (SBM). To determine whether three Aedesmosquito species have similar plasticity in SSD, attain sexual dimorphism through similar strategies, and whether SSD and SBM are associated. Aedes albopictus, Aedes aegypti, and Aedes triseriatus (Diptera: Culicidae). In four different food availability environments, we quantified plastic responses of relative growth rate (RGR), development time, and adult body size in individually reared males and females. Food availability affected RGR differently for the sexes for all three species. The RGR of males and females differed significantly in the 0.1 g/L food treatment. This difference did not account for observed SSD. Food levels over which the largest changes in RGR were observed differed among the species. Male and female adult mass and development time were jointly affected by food availability in a pattern that differed among the three species, so that degree of SSD and SBM changed differentially with food availability for all three species. Development time was generally less sexually dimorphic than mass, particularly in A. albopictus. At lower food levels, A. aegypti and A. triseriatus had accentuated dimorphism in development time. These results, combined with our knowledge of mosquito life history, suggest that a direct benefit of SBM is improbable for mosquitoes and that the observed intersexual differences in development time are more likely byproducts of selection for SSD.

Aedesmosquitoes are highly invasive and can survive almost any climatic conditions. They transmit a number of major world's deadly diseases. Therefore, a study was undertaken during December 2010 to evaluate the entomo-epidemiological risk of Aedesmosquito borne diseases (VBD) in Mumbai international seaport areas to minimize potential global health risks and prevent introduction of new VBD in India. Surveys were undertaken in operational and residential areas of Mumbai Port Trust (MPT). Al...

The density of Aedes sp in a region has critical influence on the high incidence of dengue. This aims of study was to determine the effect of modification of ovitrap and attractant substances on the density of Aedes sp mosquitoes in endemic areas of Makassar. The research type was experimental with pretest-posttest Randomized Control Design. Samples are eggs and larvae present in the household who was arrested with ovitrap and attractants. The results of the study reported that mosquito densi...

An in depth understanding of mosquito breeding biology and factors regulating population sizes is fundamental for vector population control. This paper presents results from a survey of mosquito breeding habitats and the efficacy of neem chippings as a potential larvicide that can be integrated in mosquito control on Nyabondo Plateau in western Kenya. Six main mosquito habitat types namely artificial ponds, abandoned fish ponds, active fish ponds, open drains, temporary pools and swamps were found in Nyabondo. Early anopheline instars were mainly recovered from temporary pools, artificial ponds and abandoned fish ponds. The mosquitoes collected were Anopheles gambiae sensu lato (35%), An. coustani (46%) and Culex spp (19%). Both early and late instar larvae of anopheline and culicine mosquitoes were more abundant in the controls than in the Bti and neem treated habitats. Within treated habitats, early instar anopheline mosquitoes were recovered more from habitats provided with neem and fish compared to Bti treated habitats. All treated habitats recorded higher numbers of early instar larvae than late instars or pupae, indicating that gravid female mosquitoes still oviposited within treated habitats. Neem chippings are a good tool for mosquitolarval source management under field conditions. However, more research needs to be done to quantify the contribution of this tool to the overall mosquito borne disease transmission.

Full Text Available Aedes albopictus, a vector of Dengue and Chikungunya viruses, is a robust invasive species in both tropical and temperate environments. MicroRNAs (miRNAs regulate gene expression and biological processes including embryonic development, innate immunity and infection. While a number of miRNAs have been discovered in some mosquitoes, no comprehensive effort has been made to characterize them from different developmental stages from a single species. Systematic analysis of miRNAs in Ae. albopictus will improve our understanding of its basic biology and inform novel strategies to prevent virus transmission. Between 10-14 million Illumina sequencing reads per sample were obtained from embryos, larvae, pupae, adult males, sugar-fed and blood-fed adult females. A total of 119 miRNA genes represented by 215 miRNA or miRNA star (miRNA* sequences were identified, 15 of which are novel. Eleven, two, and two of the newly-discovered miRNA genes appear specific to Aedes, Culicinae, and Culicidae, respectively. A number of miRNAs accumulate predominantly in one or two developmental stages and the large number that showed differences in abundance following a blood meal likely are important in blood-induced mosquito biology. Gene Ontology (GO analysis of the targets of all Ae. albopictus miRNAs provides a useful starting point for the study of their functions in mosquitoes. This study is the first systematic analysis of miRNAs based on deep-sequencing of small RNA samples of all developmental stages of a mosquito species. A number of miRNAs are related to specific physiological states, most notably, pre- and post-blood feeding. The distribution of lineage-specific miRNAs is consistent with mosquito phylogeny and the presence of a number of Aedes-specific miRNAs likely reflects the divergence between the Aedes and Culex genera.

Aedes albopictus, a vector of Dengue and Chikungunya viruses, is a robust invasive species in both tropical and temperate environments. MicroRNAs (miRNAs) regulate gene expression and biological processes including embryonic development, innate immunity and infection. While a number of miRNAs have been discovered in some mosquitoes, no comprehensive effort has been made to characterize them from different developmental stages from a single species. Systematic analysis of miRNAs in Ae. albopictus will improve our understanding of its basic biology and inform novel strategies to prevent virus transmission. Between 10-14 million Illumina sequencing reads per sample were obtained from embryos, larvae, pupae, adult males, sugar-fed and blood-fed adult females. A total of 119 miRNA genes represented by 215 miRNA or miRNA star (miRNA*) sequences were identified, 15 of which are novel. Eleven, two, and two of the newly-discovered miRNA genes appear specific to Aedes, Culicinae, and Culicidae, respectively. A number of miRNAs accumulate predominantly in one or two developmental stages and the large number that showed differences in abundance following a blood meal likely are important in blood-induced mosquito biology. Gene Ontology (GO) analysis of the targets of all Ae. albopictus miRNAs provides a useful starting point for the study of their functions in mosquitoes. This study is the first systematic analysis of miRNAs based on deep-sequencing of small RNA samples of all developmental stages of a mosquito species. A number of miRNAs are related to specific physiological states, most notably, pre- and post-blood feeding. The distribution of lineage-specific miRNAs is consistent with mosquito phylogeny and the presence of a number of Aedes-specific miRNAs likely reflects the divergence between the Aedes and Culex genera.

Full Text Available Abstract Background Arthropod-borne viruses (arboviruses can persistently infect and cause limited damage to mosquito vectors. RNA interference (RNAi is a mosquito antiviral response important in restricting RNA virus replication and has been shown to be active against some arboviruses. The goal of this study was to use a recombinant Sindbis virus (SINV; family Togaviridae; genus Alphavirus that expresses B2 protein of Flock House virus (FHV; family Nodaviridae; genus Alphanodavirus, a protein that inhibits RNAi, to determine the effects of linking arbovirus infection with RNAi inhibition. Results B2 protein expression from SINV (TE/3'2J inhibited the accumulation of non-specific small RNAs in Aedes aegypti mosquito cell culture and virus-specific small RNAs both in infected cell culture and Ae. aegypti mosquitoes. More viral genomic and subgenomic RNA accumulated in cells and mosquitoes infected with TE/3'2J virus expressing B2 (TE/3'2J/B2 compared to TE/3'2J and TE/3'2J virus expressing GFP. TE/3'2J/B2 exhibited increased infection rates, dissemination rates, and infectious virus titers in mosquitoes following oral bloodmeal. Following infectious oral bloodmeal, significantly more mosquitoes died when TE/3'2J/B2 was ingested. The virus was 100% lethal following intrathoracic inoculation of multiple mosquito species and lethality was dose-dependent in Ae. aegypti. Conclusion We show that RNAi is active in Ae. aegypti cell culture and that B2 protein inhibits RNAi in mosquito cells when expressed by a recombinant SINV. Also, SINV more efficiently replicates in mosquito cells when RNAi is inhibited. Finally, TE/3'2J/B2 kills mosquitoes in a dose-dependent manner independent of infection route and mosquito species.

Aedes aegypti is the main vector of dengue disease in many tropical and sub-tropical countries. Dengue became major public concern in these countries due to the unavailability of vaccine or drugs for dengue disease in the market. Hence, the only way to control the spread of DF and DHF is by controlling the vectors carrying the disease, for instance with fumigation, temephos or genetic manipulation. Many previous studies conclude that Aedes aegypti may develop resistance to many kind of insecticide, including temephos. Mathematical model for transmission of temephos resistance in Aedes aegypti population is discussed in this paper. Nontrivial equilibrium point of the system and the corresponding existence are shown analytically. The model analysis have shown epidemiological trends condition that permits the coexistence of nontrivial equilibrium is given analytically. Numerical results are given to show parameter sensitivity and some cases of worsening effect values for illustrating possible conditions in the field.

Mosquitoes are one of the most important vectors of human disease. The ability of mosquitoes to transmit disease is dependent on the age structure of the population, as mosquitoes must survive long enough for the parasites to complete their development and infect another human. Age could have additional effects due to mortality rates and vector competence changing as mosquitoes senesce, but these are comparatively poorly understood. We have investigated these factors using the mosquitoAedes aegypti and the filarial nematode Brugia malayi. Rather than observing any effects of immune senescence, we found that older mosquitoes were more resistant, but this only occurred if they had previously been maintained on a nutrient-poor diet of fructose. Constant blood feeding reversed this decline in vector competence, meaning that the number of parasites remained relatively unchanged as mosquitoes aged. Old females that had been maintained on fructose also experienced a sharp spike in mortality after an infected blood meal ("refeeding syndrome") and few survived long enough for the parasite to develop. Again, this effect was prevented by frequent blood meals. Our results indicate that old mosquitoes may be inefficient vectors due to low vector competence and high mortality, but that frequent blood meals can prevent these effects of age.

Full Text Available INTRODUCTION: Dengue is one of the most widespread mosquito-borne diseases in the world. The causative agent, dengue virus (DENV, is primarily transmitted by the mosquitoAedes aegypti, a species that has proved difficult to control using conventional methods. The discovery that A. aegypti transinfected with the wMel strain of Wolbachia showed limited DENV replication led to trial field releases of these mosquitoes in Cairns, Australia as a biocontrol strategy for the virus. METHODOLOGY/PRINCIPAL FINDINGS: Field collected wMel mosquitoes that were challenged with three DENV serotypes displayed limited rates of body infection, viral replication and dissemination to the head compared to uninfected controls. Rates of dengue infection, replication and dissemination in field wMel mosquitoes were similar to those observed in the original transinfected wMel line that had been maintained in the laboratory. We found that wMel was distributed in similar body tissues in field mosquitoes as in laboratory ones, but, at seven days following blood-feeding, wMel densities increased to a greater extent in field mosquitoes. CONCLUSIONS/SIGNIFICANCE: Our results indicate that virus-blocking is likely to persist in Wolbachia-infected mosquitoes after their release and establishment in wild populations, suggesting that Wolbachia biocontrol may be a successful strategy for reducing dengue transmission in the field.

The recent emergence of both chikungunya and Zika viruses in the Americas has significantly expanded their distribution and has thus increased the possibility that individuals may become infected by more than one Aedes aegypti-borne virus at a time. Recent clinical data support an increase in the frequency of coinfection in human patients, raising the likelihood that mosquitoes could be exposed to multiple arboviruses during one feeding episode. The impact of coinfection on the ability of relevant vector species to transmit any of these viruses (that is, their vector competence) has not been determined. Thus, we here expose Ae. aegypti mosquitoes to chikungunya, dengue-2 or Zika viruses, both individually and as double and triple infections. Our results show that these mosquitoes can be infected with and can transmit all combinations of these viruses simultaneously. Importantly, infection, dissemination and transmission rates in mosquitoes are only mildly affected by coinfection. PMID:28524874

This video protocol demonstrates an effective technique to knockdown a particular gene in an insect and conduct a novel bioassay to measure excretion rate. This method can be used to obtain a better understanding of the process of diuresis in insects and is especially useful in the study of diuresis in blood-feeding arthropods that are able to take up huge amounts of liquid in a single blood meal. This RNAi-mediated gene knockdown combined with an in vivo diuresis assay was developed by the Hansen lab to study the effects of RNAi-mediated knockdown of aquaporin genes on Aedes aegypti mosquito diuresis. The protocol is setup in two parts: the first demonstration illustrates how to construct a simple mosquito injection device and how to prepare and inject dsRNA into the thorax of mosquitoes for RNAi-mediated gene knockdown. The second demonstration illustrates how to determine excretion rates in mosquitoes using an in vivo bioassay.

Full Text Available A cross-sectional study was conducted in urban community of Thanlyin Township, Yangon Region during 2014 to determine Aedeslarval indices and the occurrence of Dengue Haemorrhagic Fever (DHF within past one year. A total of 327 households and 1491 members were included in the study. Aedeslarval indices detected in this study were 25.7% for house index, 15.5% for container index and 48.0% for Breteau index. The occurrence of DHF among households and family members were 2.1% (95% CI: 0.9%, 4.4% and 0.6% (95% CI: 0.3%, 1.1%, respectively. The occurrence was highest among 5 to 14 years age-group. No case was reported among persons with equal or more than 60 years of age. Mortality and case fatality rates were 0% during study period. Larval positivity among households was significantly related to sufficiency of family income and number of water container they have. Surveillance and control procedures for both DHF and vector should be intensified in urban area. Awareness and participation of the community in prevention and control of DHF should also be raised. Socioeconomic status as well as proper water supply and storage should be improved in urban area.

Distributions of mosquito larvae likely are a consequence of multiple factors, although two commonly studied factors (quality of the larval environment and the terrestrial matrix in which these habitats reside) have rarely and simultaneously been varied in the field to understand...

ABSTRACT. The Asian exotic mosquitoAedes japonicus was 1st collected in Minnesota in 2007 and was well established in parts of the Metropolitan Mosquito Control District (MMCD) by 2008. Surveillance strategies were devised for 2009 to track the expansion of its range through MMCD and to direct control efforts. Sampling of larvae from container and tire habitats was the primary method used to document Ae. japonicus presence, but larvae were found in other habitats as well. Adult Ae. japonicus were collected by vacuum aspirator, gravid trap, and New Jersey trap but not by CO2-baited Centers for Disease Control and Prevention light trap. Aedes japonicus were collected from each of the 7 counties surveyed; in 5 of the counties for the 1st time in 2009. Preliminary findings suggest that a control strategy involving intensive source reduction can reduce Ae. japonicus populations.

The mosquitoAedes aegypti transmits viruses that cause diseases such as dengue, chikungunya and zika. Measures are taken to control the mosquito since these infectious diseases represent a significant health problem. This is the case on the island of Saba, a Dutch Caribbean island. In order to

Since 2005, Aedes albopictus mosquitoes have been detected in companies in the Netherlands that import ornamental plants from China. To assess the risks of dengue transmission, a study was carried out in 48 persons who are professionally exposed to these mosquitoes. No evidence for

The mosquitoAedes aegypti transmits viruses that cause diseases such as dengue, chikungunya and zika. Measures are taken to control the mosquito since these infectious diseases represent a significant health problem. This is the case on the island of Saba, a Dutch Caribbean island. In order to figh

Since 2005, Aedes albopictus mosquitoes have been detected in companies in the Netherlands that import ornamental plants from China. To assess the risks of dengue transmission, a study was carried out in 48 persons who are professionally exposed to these mosquitoes. No evidence for non-travel-relate

Since 2005, Aedes albopictus mosquitoes have been detected in companies in the Netherlands that import ornamental plants from China. To assess the risks of dengue transmission, a study was carried out in 48 persons who are professionally exposed to these mosquitoes. No evidence for non-travel-relate

Ribonucleic acid interference (RNAi) is a powerful approach for elucidating gene functions in a variety of organisms, including mosquitoes and many other insects. Little has been done, however, to harness this approach in order to control adult and larvalmosquitoes. Juvenile hormone (JH) plays a pi...

Objective: To determine the effects of environmental parameters of larval habitats on distribution and abundance of anopheline mosquitoes in Rudan county of Iran. Methods: This cross-sectional study was conducted during the mosquito breeding season from February 2010 to October 2011. The anopheline larvae were collected using the standard dipping method. The specimens were identified using a morphological-based key. Simultaneously with larval collection, environmental parameters of the larval habitats including water current and turbidity, sunlight situation, and substrate type of habitats were recorded. Water samples were taken from breeding sites during larval collection. Before collection of samples, the water temperature was measured. The water samples were analysed for turbidity, conductivity, total alkalinity, total dissolved solid, pH and ions including chloride, sulphate, calcium, and magnesium. Statistical correlation analysis and ANOVA test were used to analyze the association between environmental parameters and larvalmosquito abundance. Results: In total 2 973 larvae of the genus Anopheles were collected from 25 larval habitats and identified using morphological characters. They comprised of six species:An. dthali turkhudi (3.30%), and An. apoci (1.14%). The most abundant species was An. dthali which were collected from all of the study areas. Larvae of two malaria vectors, An. dthali and An. stephensi, co-existed and collected in a wide range of habitats with different physico-chemical parameters. The most common larval habitats were man-made sites such as sand mining pools with clean and still water. The anopheline mosquitoes also preferred permanent habitats in sunlight with sandy substrates. The results indicated that there was a significant relationship between mean physico-chemical parameters such as water temperature, conductivity, total alkalinity, sulphate, chloride, and mosquito distribution and abundance. Conclusions: The results of this

The occurrence of the Asian tiger mosquito, Aedes albopictus , has been reported in the Mexican states of Tamaulipas, Nuevo León, Coahuila (northeastern), Veracruz, Chiapas, Quintana Roo (southeastern), Morelos, San Luis Potosí (middle), and Sinaloa (northwestern). In April and September 2012, Ae. albopictus was collected in a variety of habitats and landing/biting on the collecting personnel in 12 counties of Hidalgo state (middle). This is the first record of the occurrence of this species in Hidalgo state.

The value of using plant volatiles as attractants for trapping and spatial repellents to protect hosts against mosquitoes has been widely recognized. The current study characterized behavioral responses of Aedes albopictus (Skuse) (Diptera: Culicidae) to different concentrations, ranging from 6 to 96%, of several common floral odorous compounds, including linalool, geraniol, citronellal, eugenol, anisaldehyde, and citral, using a wind tunnel olfactometer system. The results indicated that fem...

.... The mosquitoes of major concern in Malaysia belong to the genera Culex, Aedes, and Armigeres. Objective: To study the host-seeking behavior of four Malaysian mosquitoes commonly found in urban residential areas in Kuala Lumpur...

Interventions targeting adult mosquitoes are used to combat transmission of vector-borne diseases, including dengue. Without available vaccines, targeting the primary vector, Aedes aegypti, is essential to prevent transmission. Older mosquitoes (>/='7 days) are of greatest epidemiological significan...

Field experiments were conducted in central Kenya, to study the effect of ammonium sulphate fertiliser ((NH(4))(2)SO(4)) on mosquitolarval populations in rice fields. The experiments used a complete randomised block design having four blocks with two experimental ponds per block, and the fertili......Field experiments were conducted in central Kenya, to study the effect of ammonium sulphate fertiliser ((NH(4))(2)SO(4)) on mosquitolarval populations in rice fields. The experiments used a complete randomised block design having four blocks with two experimental ponds per block...... populations of An. arabiensis (Pmosquitoes (P... in rice fields, thereby making them visually more attractive for egg-laying by An. arabiensis and culicine mosquitoes....

Aedes aegypti L. (Diptera: Culicidae) is a major vector of yellow fever, dengue, and chikungunya viruses throughout tropical and subtropical areas of the world. Although the southernmost part of Mauritania along the Senegal river has long been recognized at risk of yellow fever transmission, Aedes spp. mosquitoes had never been reported northwards in Mauritania. Here, we report the first observation of Aedes aegypti aegypti (L.) and Aedes (Ochlerotatus) caspius (Pallas, 1771) in the capital city, Nouakchott. We describe the development sites in which larvae of the two species were found, drawing attention to the risk for emergence of arbovirus transmission in the city.

Full Text Available In Brazil, the entomological surveillance of Aedes (Stegomyia aegypti is performed by government-mandated larval surveys. In this study, the sensitivities of an adult sticky trap and traditional surveillance methodologies were compared. The study was performed over a 12-week period in a residential neighbourhood of the municipality of Pedro Leopoldo, state of Minas Gerais, Brazil. An ovitrap and a MosquiTRAP were placed at opposite ends of each neighbourhood block (60 traps in total and inspections were performed weekly. The study revealed significant correlations of moderate strength between the larval survey, ovitrap and MosquiTRAP measurements. A positive relationship was observed between temperature, adult capture measurements and egg collections, whereas precipitation and frequency of rainy days exhibited a negative relationship.

In Brazil, the entomological surveillance of Aedes (Stegomyia) aegypti is performed by government-mandated larval surveys. In this study, the sensitivities of an adult sticky trap and traditional surveillance methodologies were compared. The study was performed over a 12-week period in a residential neighbourhood of the municipality of Pedro Leopoldo, state of Minas Gerais, Brazil. An ovitrap and a MosquiTRAP were placed at opposite ends of each neighbourhood block (60 traps in total) and inspections were performed weekly. The study revealed significant correlations of moderate strength between the larval survey, ovitrap and MosquiTRAP measurements. A positive relationship was observed between temperature, adult capture measurements and egg collections, whereas precipitation and frequency of rainy days exhibited a negative relationship.

In Brazil, the entomological surveillance of Aedes (Stegomyia) aegypti is performed by government-mandated larval surveys. In this study, the sensitivities of an adult sticky trap and traditional surveillance methodologies were compared. The study was performed over a 12-week period in a residential neighbourhood of the municipality of Pedro Leopoldo, state of Minas Gerais, Brazil. An ovitrap and a MosquiTRAP were placed at opposite ends of each neighbourhood block (60 traps in total) and inspections were performed weekly. The study revealed significant correlations of moderate strength between the larval survey, ovitrap and MosquiTRAP measurements. A positive relationship was observed between temperature, adult capture measurements and egg collections, whereas precipitation and frequency of rainy days exhibited a negative relationship. PMID:24402144

Full Text Available Abstract Background Aedes aegypti and Aedes albopictus are both vectors of chikungunya virus (CHIKV. The two Aedes species co-exist in the Indian Ocean region and were involved in the 2005-2006 CHIKV outbreaks. In the Reunion Island, a single mutation in the viral envelope has been selected that leads to high levels of replication in Ae. albopictus, and a short extrinsic incubation period as the virus could be found in saliva as early as two days after infection. An important question is whether this variant is associated with adverse effects impacting some mosquito life-history traits such as survival and reproduction. Results We performed experimental infections using three mosquito strains of Ae. aegypti Mayotte and Ae. albopictus (Mayotte and Reunion, and two CHIKV strains (E1-226A and E1-226V. Ae. aegypti Mayotte were similarly susceptible to both viral strains, whereas Ae. albopictus Mayotte and Ae. albopictus Reunion were more susceptible to CHIKV E1-226V than to E1-226A. In terms of life-history traits measured by examining mosquito survival and reproduction, we found that: (1 differences were observed between responses of mosquito species to the two viruses, (2 CHIKV infection only affected significantly some life-history traits of Ae. albopictus Reunion and not of the other two mosquito strains, and (3 CHIKV reduced the lifespan of Ae. albopictus Reunion and shortened the time before egg laying. Conclusion We demonstrated that CHIKV only reduces the survival of Ae. albopictus from the Reunion Island. By laying eggs just before death, reproduction of Ae. albopictus from the Reunion Island is not reduced since other parameters characterizing oviposition and hatching were not affected.

Aedes aegypti and Aedes albopictus are both vectors of chikungunya virus (CHIKV). The two Aedes species co-exist in the Indian Ocean region and were involved in the 2005-2006 CHIKV outbreaks. In the Reunion Island, a single mutation in the viral envelope has been selected that leads to high levels of replication in Ae. albopictus, and a short extrinsic incubation period as the virus could be found in saliva as early as two days after infection. An important question is whether this variant is associated with adverse effects impacting some mosquito life-history traits such as survival and reproduction. We performed experimental infections using three mosquito strains of Ae. aegypti Mayotte and Ae. albopictus (Mayotte and Reunion), and two CHIKV strains (E1-226A and E1-226V). Ae. aegypti Mayotte were similarly susceptible to both viral strains, whereas Ae. albopictus Mayotte and Ae. albopictus Reunion were more susceptible to CHIKV E1-226V than to E1-226A. In terms of life-history traits measured by examining mosquito survival and reproduction, we found that: (1) differences were observed between responses of mosquito species to the two viruses, (2) CHIKV infection only affected significantly some life-history traits of Ae. albopictus Reunion and not of the other two mosquito strains, and (3) CHIKV reduced the lifespan of Ae. albopictus Reunion and shortened the time before egg laying. We demonstrated that CHIKV only reduces the survival of Ae. albopictus from the Reunion Island. By laying eggs just before death, reproduction of Ae. albopictus from the Reunion Island is not reduced since other parameters characterizing oviposition and hatching were not affected.

The presence of Aedes albopictus (Skuse) in the Torres Strait of northern Australia increases the potential for colonization and establishment on the mainland. However, there is a possibility that native species that occupy the same habitats may influence the population performance of Ae. albopictus, potentially affecting the establishment of this species in Australia. Cohabitation experiments were performed with the endemic Aedes notoscriptus (Skuse), which has been found occupying the same larval habitats as Ae. albopictus in the Torres Strait and is the most widespread container-inhabiting Aedes species in Australia. The influence of environmental factors and cohabitation between the two species was examined using different climates, food resource levels, food resource types, and species densities. Survivorship proportions and a population performance index (λ') were calculated and compared. The consequences of increased Ae. notoscriptus densities were reduced survivorship and λ' for Ae. albopictus. Despite this, the mean λ' of Ae. albopictus and Ae. notoscriptus was consistently ≥ 1.06, indicating both species could increase under all conditions, potentially due to increasing conspecific densities negatively affecting Ae. notoscriptus. The outcomes from this study suggest that the preexisting presence of Ae. notoscriptus may not prevent the establishment of Ae. albopictus in Australia.

Full Text Available Objective. To investigate if family head genre-associated education is related to the risk of domiciliary Aedes aegypti larval breeding in a dengue-endemic village of Southern Mexico. Material and Methods. A family head was considered to have a low education level if he/she had not completed elementary school. To estimate larval breeding risk within each household, a three-category Maya index was constructed using a weighted estimation of controllable and disposable domestic water containers. A socio-economic index was constructed based on household construction characteristics. Results. Low-level education of either family head was associated to higher larval breeding risk. Households with low-educated mothers had more larval breeding containers. These associations persisted after adjusting for household socio-economic level. Conclusions. These results indicate that households with female family heads with low education levels accumulate more containers that favor Ae. aegypti breeding, and that education campaigns for dengue control should be addressed to this part of the population.

Repellent efficacy of the Off! Clip-on Mosquito Repellent device (S. C. Johnson and Son, Inc., Racine, WI) containing Metofluthrin was evaluated on six human volunteers against the container-breeding mosquitoAedes albopictus (Skuse) and the salt marsh mosquitoAedes taeniorhynchus (Wiedemann) at two field locations in northeastern Florida. The device repelled mosquitoes by releasing a vaporized form of the pyrethroid insecticide metofluthrin ([AI] 31.2%) and provided 70% protection from Ae. albopictus bites for > 3 h. For the second field trial, a repellent device that was used in the first trial was tested after being open for >1 wk. This device provided 79% protection from Ae. taeniorhynchus bites for 3 h. Our field results showed that the repellent device was 70 and 79% effective at repelling Ae. albopictus and Ae. taeniorhynchus from human test subjects in both field locations in northeastern Florida.

Mosquito habitat-association studies are an important basis for disease control programmes and/or vector distribution models. However, studies do not explicitly account for incomplete detection during larval presence and abundance surveys, with potential for significant biases because of environmental influences on larval behaviour and sampling efficiency. Data were used from a dip-sampling study for Anopheles larvae in Ethiopia to evaluate the effect of six factors previously associated with larval sampling (riparian vegetation, direct sunshine, algae, water depth, pH and temperature) on larval presence and detectability. Comparisons were made between: (i) a presence-absence logistic regression where samples were pooled at the site level and detectability ignored, (ii) a success versus trials binomial model, and (iii) a presence-detection mixture model that separately estimated presence and detection, and fitted different explanatory variables to these estimations. Riparian vegetation was consistently highlighted as important, strongly suggesting it explains larval presence (-). However, depending on how larval detectability was estimated, the other factors showed large variations in their statistical importance. The presence-detection mixture model provided strong evidence that larval detectability was influenced by sunshine and water temperature (+), with weaker evidence for algae (+) and water depth (-). For larval presence, there was also some evidence that water depth (-) and pH (+) influenced site occupation. The number of dip-samples needed to determine if larvae were likely present at a site was condition dependent: with sunshine and warm water requiring only two dips, while cooler water and cloud cover required 11. Environmental factors influence true larval presence and larval detectability differentially when sampling in field conditions. Researchers need to be more aware of the limitations and possible biases in different analytical approaches used to

Sex determination in the mosquitoAedes aegypti is governed by a dominant male-determining factor (M factor) located within a Y chromosome-like region called the M locus. Here, we show that an M-locus gene, Nix, functions as an M factor in A. aegypti. Nix exhibits persistent M linkage and early embryonic expression, two characteristics required of an M factor. Nix knockout with clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 resulted in largely feminized genetic males and the production of female isoforms of two key regulators of sexual differentiation: doublesex and fruitless. Ectopic expression of Nix resulted in genetic females with nearly complete male genitalia. Thus, Nix is both required and sufficient to initiate male development. This study provides a foundation for mosquito control strategies that convert female mosquitoes into harmless males.

In repeated behaviours such as those of feeding and reproduction, past experiences can inform future behaviour. By altering their behaviour in response to environmental stimuli, insects in highly variable landscapes can tailor their behaviour to their particular environment. In particular, female mosquitoes may benefit from plasticity in their choice of egg-laying site as these sites are often temporally variable and clustered. The opportunity to adapt egg-laying behaviour to past experience also exists for mosquito populations as females typically lay eggs multiple times throughout their lives. Whether experience and age affect egg-laying (or oviposition) behaviour in the mosquito Stegomyia aegypti (=Aedes aegypti) (Diptera: Culicidae) was assessed using a wind tunnel. Initially, gravid mosquitoes were provided with a cup containing either repellent or well water. After ovipositing in these cups, the mosquitoes were blood-fed and introduced into a wind tunnel. In this wind tunnel, an oviposition cup containing repellent was placed in the immediate vicinity of the gravid mosquitoes. A cup containing well water was placed at the opposite end of the tunnel so that if the females flew across the chamber, they encountered the well water cup, in which they readily laid eggs. Mosquitoes previously exposed to repellent cups became significantly more likely to later lay eggs in repellent cups, suggesting that previous experience with suboptimal oviposition sites informs mosquitoes of the characteristics of nearby oviposition sites. These results provide further evidence that mosquitoes modify behaviour in response to environmental information and are demonstrated in a vector species in which behavioural plasticity may be ecologically and epidemiologically meaningful.

Full Text Available Objective: To determine the infestation profile by performing ovitrap surveillance in four types of residential areas in central zone of Shah Alam, Selangor. Methods: 600 ovitraps were exposed for 4 days in intradomiciliary environments within 20 residential areas including independent houses, inter-connected houses, commercial buildings and mixed houses areas. The ecological conditions of the surrounding environment were observed and recorded. The distributions and abundances of the Aedesmosquitoes were presented in terms of positive ovitrap index (POI and mean eggs per trap (MET. One-way ANOVA test was used to determine the association between the type of residential areas and the number of ovitrap with eggs and the egg count. Results: The result implied that Seksyen 7 has the highest reading of both ovitrap index and egg count (POI = 86.21%, MET = 41.68 eggs. There was no significant difference in the POI and MET of Aedesmosquitoes between the type of housing areas (P > 0.05. The surrounding environmental conditions were believed to be the main influence of the abundance and distribution of the Aedesmosquitoes in Shah Alam. Unhygienic conditions were observed in all type of residential areas. These included the abandoned kitchen utensils and flower pots stored at the backyard which lead to collected stagnant water that later served as the favourable places for female Aedes to oviposit. Conclusions: The key to effectively control the dengue outbreak is to have an advance understanding to all the high potential transmission areas or habitat which are generally the areas with the densest human and vector population.

Full Text Available BACKGROUND: The mosquitoAedes aegypti was recently transinfected with a life-shortening strain of the endosymbiont Wolbachia pipientis (wMelPop as the first step in developing a biocontrol strategy for dengue virus transmission. In addition to life-shortening, the wMelPop-infected mosquitoes also exhibit increased daytime activity and metabolic rates. Here we sought to quantify the blood-feeding behaviour of Wolbachia-infected females as an indicator of any virulence or energetic drain associated with Wolbachia infection. METHODOLOGY/PRINCIPAL FINDINGS: In a series of blood-feeding trials in response to humans, we have shown that Wolbachia-infected mosquitoes do not differ in their response time to humans, but that as they age they obtain fewer and smaller blood meals than Wolbachia-uninfected controls. Lastly, we observed a behavioural characteristic in the Wolbachia infected mosquitoes best described as a "bendy" proboscis that may explain the decreased biting success. CONCLUSIONS/SIGNIFICANCE: Taken together the evidence suggests that wMelPop infection may be causing tissue damage in a manner that intensifies with mosquito age and that leads to reduced blood-feeding success. These behavioural changes require further investigation with respect to a possible physiological mechanism and their role in vectorial capacity of the insect. The selective decrease of feeding success in older mosquitoes may act synergistically with other Wolbachia-associated traits including life-shortening and viral protection in biocontrol strategies.

Dengue represents a serious threat to human health, with billions of people living at risk of the disease. Wolbachia pipientis is a bacterial endosymbiont common to many insect species. Wolbachia transinfections in mosquito disease vectors have great value for disease control given the bacterium's ability to spread into wild mosquito populations, and to interfere with infections of pathogens, such as dengue virus. Aedes fluviatilis is a mosquito with a widespread distribution in Latin America, but its status as a dengue vector has not been clarified. Ae. fluviatilis is also naturally infected by the wFlu Wolbachia strain, which has been demonstrated to enhance infection with the avian malarial parasite Plasmodium gallinaceum. We performed experimental infections of Ae. fluviatilis with DENV-2 and DENV-3 isolates from Brazil via injection or oral feeding to provide insight into its competence for the virus. We also examined the effect of the native Wolbachia infection on the virus using a mosquito line where the wFlu infection had been cleared by antibiotic treatment. Through RT-qPCR, we observed that Ae. fluviatilis could become infected with both viruses via either method of infection, although at a lower rate than Aedes aegypti, the primary dengue vector. We then detected DENV-2 and DENV-3 in the saliva of injected mosquitoes, and observed that injection of DENV-3-infected saliva produced subsequent infections in naïve Ae. aegypti. However, across our data we observed no difference in prevalence of infection and viral load between Wolbachia-infected and -uninfected mosquitoes, suggesting that there is no effect of wFlu on dengue virus. Our results highlight that Ae. fluviatilis could potentially serve as a dengue vector under the right circumstances, although further testing is required to determine if this occurs in the field.

Ovitrap surveillance was initiated for eight continuous weeks to determine the distribution and abundance of Aedes sp. mosquitoes in the University of Malaya campus, Kuala Lumpur, and the impact of meteorological conditions on the Aedes populations. Two study areas within the campus were selected: Varsity Lake and Seventh Residential College. The abundance of Aedes populations in Varsity Lake was indicated by ovitrap index (OI) which ranged from 60.00%-90.00%. The mean number of larvae per ovitrap of Aedes albopictus in Varsity Lake ranged from 11.23+/-2.42-43.80+/-6.22. On the other hand, the outdoor OI for Seventh Residential College ranged from 73.33%-93.33%, respectively, while the mean number larvae per ovitrap for this area ranged from 19.33+/-4.55-35.27+/-5.46, respectively. In addition, the indoor OI of Seventh Residential College ranged from 0.00%-30.00%, while the mean number of larvae per ovitrap for Ae. albopictus ranged from 0-5.90+/-3.55. There was no significant difference (p>0.05) of Ae. albopictus population between Varsity Lake and Seventh Residential College. The studies showed a correlation between OI and mean number of larvae per ovitrap for outdoor Ae. albopictus populations in Varsity Lake and Seventh Residential College (r=0.794). There was also a correlation between the mean larvae number per ovitrap of Ae. albopictus obtained from eight weeks indoor ovitrap surveillance in Seventh Residential College with rainfall (r=0.584). However, there was no correlation between the mean larvae number per ovitrap of Ae. albopictus in both study areas with temperature and relative humidity. Aedes aegypti mosquitoes were found neither indoor nor outdoor in both study areas. This study indicated that the principal dengue vector in the university campus was most likely Ae. albopictus.

Full Text Available We aimed to extract the ingredients from leaves of Gossypium hirsutum (Bt cotton using different solvents and evaluate for potential use to control different larval stages of mosquito species, Aedes aegypti and Anopheles stephensi.Qualitative and quantitative estimation of ingredients from Go. hirsutum (Bt plant extract was carried out and their inhibitory action against mosquito larvae was determined using mosquito larvicidal assay.LC50 values of water, ethanol, ethyl acetate and hexane extracts for Ae. aegypti were 211.73±21.49, 241.64±19.92, 358.07±32.43, 401.03±36.19 and 232.56±26.00, 298.54±21.78, 366.50±30.59, 387.19±31.82 for 4(th instar of An. stephensi, respectively. The water extract displayed lowest LC50 value followed by ethanol, ethyl acetate and hexane. Owing to the comparatively better activity of water extract, its efficacy was further evaluated for mosquito larvicidal activity, which exhibited LC50 values of 133.95±12.79, 167.65±11.34 against 2(nd and 3(rd instars of Ae. aegypti and 145.48±11.76, 188.10±12.92 against 2(nd and 3(rd instars of An. stephensi, respectively. Crude protein from the water extract was precipitated using acetone and tested against 2(nd, 3(rd and 4(th instars of Ae. aegypti and An. stephensi. It revealed further decrease in LC50 values as 105.72±25.84, 138.23±23.18, 126.19±25.65, 134.04±04 and 137.88±17.59, 154.25±16.98 for 2(nd, 3(rd and 4(th instars of Ae. aegypti and An. stephensi, respectively.Leaves extracts of Go. hirsutum (Bt is potential mosquito larvicide and can be used as a potent alternative to chemical insecticides in integrated pest management.

Full Text Available Several important diseases are transmitted by mosquitoes. Despite of the potential of the occurrence of some mosquito-borne diseases such as West Nile, dirofilariasis and malaria in the region, there is no recent study of mosquitoes in West Azerbaijan Province. The aim of this investigation was to study the fauna, composition and distribution of mosquitoes and the characteristics of their larval habitats in this province.Larvae and adult collections were carried out from different habitats using the standard methods in twenty five localities of seven counties across West Azerbaijan Province.Overall, 1569 mosquitoes including 1336 larvae and 233 adults were collected from 25 localities. The details of geographical properties were recorded. Five genera along with 12 species were collected and identified including: Anopheles claviger, An. maculipennis s.l., An. superpictus, Culex pipiens, Cx. theileri, Cx. modestus, Cx. hortensis, Cx. mimeticus, Culiseta Longiareolata, Ochlerotatus caspius s.l., Oc. geniculatus and Uranotaenia unguiculata. This is the first record of Oc. geniculatus in the province.Due to the geographical location of the West Azerbaijan Province, it comprises different climatic condition which provides suitable environment for the establishment of various species of mosquitoes. The solidarity geographical, cultural and territorial exchanges complicate the situation of the province and its vectors as a threat for future and probable epidemics of mosquito-borne diseases.

Full Text Available The failure of traditional vector control operations through chemical insecticides renewed interest in biological control method. In the present study Dragonfly (Brachytron pratense nymph has been proved to be a strong biocontrol agent of Anopheles suhpictus larvae in the laboratory condition. Average daily larval feeding rate of B. pratense nymph decreased when the search area was increased. Feeding rate increased when prey density was increased. In the field conditions also, B. pratense played very effective role as predator of different species of larvalmosquitoes.

Full Text Available The failure of traditional vector control operations through chemical insecticides renewed interest in biological control method. In the present study Dragonfly (Brachytron pratense nymph has been proved to be a strong biocontrol agent of Anopheles suhpictus larvae in the laboratory condition. Average daily larval feeding rate of B. pratense nymph decreased when the search area was increased. Feeding rate increased when prey density was increased. In the field conditions also, B. pratense played very effective role as predator of different species of larvalmosquitoes.

The mosquitoesAedes aegypti and Aedes albopictus are vectors of pathogenic viruses that cause major human illnesses including dengue, yellow fever and chikungunya. Both mosquito species are expanding their geographic distributions and now occur worldwide in temperate and tropical climates. Collection of eggs in oviposition traps (ovitraps) is commonly used for monitoring and surveillance of container-inhabiting Aedes populations by public health agencies charged with managing mosquito-transmitted illness. Addition of an organic infusion in these traps increases the number of eggs deposited. Gravid females are guided to ovitraps by volatile chemicals produced from the breakdown of organic matter by microbes. We previously isolated and cultured 14 species of bacteria from attractive experimental infusions, made from the senescent leaves of canebrake bamboo (Arundinaria gigantea). Cultures were grown for 24 h at 28 °C with constant shaking (120 rpm) and cell densities were determined with a hemocytometer. Behavioral responses to single bacterial isolates and to a mix of isolates at different cell densities were evaluated using two-choice sticky-screen bioassay methods with gravid Ae. aegypti and Ae. albopictus. In behavioral assays of a mix of 14 bacterial isolates, significantly greater attraction responses were exhibited by Ae. aegypti and Ae. albopictus to bacterial densities of 10(7) and 10(8) cells/mL than to the control medium. When we tested single bacterial isolates, seven isolates (B1, B2, B3, B5, B12, B13 and B14) were significantly attractive to Ae. aegypti, and six isolates (B1, B5, B7, B10, B13 and B14) significantly attracted Ae. albopictus. Among all the isolates tested at three different cell densities, bacterial isolates B1, B5, B13 and B14 were highly attractive to both Aedes species. Our results show that at specific cell densities, some bacteria significantly influence the attraction of gravid Ae. aegypti and Ae. albopictus females to

Full Text Available The mosquito nuisance and the mosquito borne diseases have become major important challenging public health problems in India especially in the fast developing city like Pondicherry urban agglomeration. The Pondicherry government has been implemented full-fledged mosquito control measures, however, dengue and chikungunya epidemics was accelerating trend in Pondicherry for the recent years, and therefore, the directorate of public health, Pondicherry was requested vector control research centre (VCRC, to conduct a mosquito control evaluation survey. A team of field staff of VCRC headed by the author, Pondicherry, have conducted a detailed reconnaissance survey for collecting the site specifications of houses and the streetwise mosquito data for analyzing the density of vector mosquitoes in the wards / blocks and delineating the areas vulnerable to disease epidemics in the urban areas. The GPS GARMIN 12 XL was used to collect the field data. The ARC GIS 10.0 software was used to map the site locations (houses with mosquito’s data. The digital map of block boundary of Pondicherry was used for mapping purpose. A systematic grid sampling was applied to conduct a rapid survey for mapping Aedes species mosquito genic condition in the urban areas and the coordinates of sites of house information with breeding habitats positive in the grid sectors was collected using GPS, and the mean value of positive habitats was analyzed by quintiles method for mapping. The four blocks were selected for Aedesmosquito survey where the mosquito problem was identified as comparatively high, four numbers of wards were selected from each block, and the 40 number of houses was selected with 100 meter interval distance for mosquito breeding survey in the domestic and peripheral domestic areas in each wards. The problematic areas were identified, highlighted and recommended for web mapping GIS for Aedesmosquito surveillance continuously for monitoring the mosquito control

The aquatic bodies designated as mosquitolarval habitats are diverse in size and species composition. The macroinvertebrate predators in these habitats are elements that influence the abundance of mosquito species, providing a basis for biological control. Assessment of species assemblage in these habitats will indicate the possible variations in the resource exploitation and trophic interactions and, therefore, can help to frame biological control strategies more appropriately. In the present study, the species composition is being investigated in five different mosquitolarval habitats at a spatial scale. A random sample of 80 each of the habitats, grouped as either small or large, was analyzed in respect to the macroinvertebrate species assemblage. The species composition in the habitats was noted to be an increasing function of habitat size (species number = 1.653 + 0.819 habitat size) and, thus, the diversity. The relative abundance of the mosquito immatures varied with the habitat, and the number of useful predator taxa was higher in the larger habitats. In the smaller habitats-plastic and earthen structures and sewage drains, the relative and absolute number of mosquito immatures per sampling unit were significantly higher than the pond and rice field habitats. This was evident in the cluster analysis where the smaller habitats were more related than the larger habitats. The principal component analysis on the species diversity yielded four and six components, respectively, for the smaller and larger habitats for explaining the observed variance of species abundance. The species composition in the habitats was consistent with the earlier findings and support that the abundance of coexisting macroinvertebrate species regulates the relative load of mosquito immatures in the habitats. The findings of this study may be further tested to deduce the relative importance of the habitats in terms of the productivity of mosquito immatures at a temporal scale.

Field experiments were conducted in central Kenya, to study the effect of ammonium sulphate fertiliser ((NH(4))(2)SO(4)) on mosquitolarval populations in rice fields. The experiments used a complete randomised block design having four blocks with two experimental ponds per block, and the fertiliser and control treatments allocated randomly among the ponds. Student's two-sample unpaired t-test was used to test for the significance of differences between the relative counts of larvae in fertiliser and control treatments. The results showed a significant overall increase in the larval populations of An. arabiensis (Price fields, thereby making them visually more attractive for egg-laying by An. arabiensis and culicine mosquitoes.

Full Text Available The Asian tiger mosquito (Aedes albopictus is an important vector for pathogens that affect human health, including the viruses that cause dengue and Chikungunya fevers. It is also one of the world's fastest-spreading invasive species. For these reasons, it is crucial to identify strategies for controlling the reproduction and spread of this mosquito. During mating, seminal fluid proteins (Sfps are transferred from male mosquitoes to females, and these Sfps modulate female behavior and physiology in ways that influence reproduction. Despite the importance of Sfps on female reproductive behavior in mosquitoes and other insects, the identity of Sfps in Ae. albopictus has not previously been reported. We used transcriptomics and proteomics to identify 198 Sfps in Ae. albopictus. We discuss possible functions of these Sfps in relation to Ae. albopictus reproduction-related biology. We additionally compare the sequences of these Sfps with proteins (including reported Sfps in several other species, including Ae. aegypti. While only 72 (36.4% of Ae. albopictus Sfps have putative orthologs in Ae. aegypti, suggesting low conservation of the complement of Sfps in these species, we find no evidence for an elevated rate of evolution or positive selection in the Sfps that are shared between the two Aedes species, suggesting high sequence conservation of those shared Sfps. Our results provide a foundation for future studies to investigate the roles of individual Sfps on feeding and reproduction in this mosquito. Functional analysis of these Sfps could inform strategies for managing the rate of pathogen transmission by Ae. albopictus.

Full Text Available In vivo targeted gene disruption is a powerful tool to study gene function. Thus far, two tools for genome editing in Aedes aegypti have been applied, zinc-finger nucleases (ZFN and transcription activator-like effector nucleases (TALEN. As a promising alternative to ZFN and TALEN, which are difficult to produce and validate using standard molecular biological techniques, the clustered regularly interspaced short palindromic repeats/CRISPR-associated sequence 9 (CRISPR/Cas9 system has recently been discovered as a "do-it-yourself" genome editing tool. Here, we describe the use of CRISPR/Cas9 in the mosquito vector, Aedes aegypti. In a transgenic mosquito line expressing both Dsred and enhanced cyan fluorescent protein (ECFP from the eye tissue-specific 3xP3 promoter in separated but tightly linked expression cassettes, we targeted the ECFP nucleotide sequence for disruption. When supplying the Cas9 enzyme and two sgRNAs targeting different regions of the ECFP gene as in vitro transcribed mRNAs for germline transformation, we recovered four different G1 pools (5.5% knockout efficiency where individuals still expressed DsRed but no longer ECFP. PCR amplification, cloning, and sequencing of PCR amplicons revealed indels in the ECFP target gene ranging from 2-27 nucleotides. These results show for the first time that CRISPR/Cas9 mediated gene editing is achievable in Ae. aegypti, paving the way for further functional genomics related studies in this mosquito species.

In vivo targeted gene disruption is a powerful tool to study gene function. Thus far, two tools for genome editing in Aedes aegypti have been applied, zinc-finger nucleases (ZFN) and transcription activator-like effector nucleases (TALEN). As a promising alternative to ZFN and TALEN, which are difficult to produce and validate using standard molecular biological techniques, the clustered regularly interspaced short palindromic repeats/CRISPR-associated sequence 9 (CRISPR/Cas9) system has recently been discovered as a "do-it-yourself" genome editing tool. Here, we describe the use of CRISPR/Cas9 in the mosquito vector, Aedes aegypti. In a transgenic mosquito line expressing both Dsred and enhanced cyan fluorescent protein (ECFP) from the eye tissue-specific 3xP3 promoter in separated but tightly linked expression cassettes, we targeted the ECFP nucleotide sequence for disruption. When supplying the Cas9 enzyme and two sgRNAs targeting different regions of the ECFP gene as in vitro transcribed mRNAs for germline transformation, we recovered four different G1 pools (5.5% knockout efficiency) where individuals still expressed DsRed but no longer ECFP. PCR amplification, cloning, and sequencing of PCR amplicons revealed indels in the ECFP target gene ranging from 2-27 nucleotides. These results show for the first time that CRISPR/Cas9 mediated gene editing is achievable in Ae. aegypti, paving the way for further functional genomics related studies in this mosquito species.

Mosquito attractants and spatial repellents hold great promise in controlling mosquito pests. In assessing the effectiveness of mosquito attractants and repellents, a good olfactometer system, and optimized testing conditions, are essential. In this research, we demonstrated the usefulness of an olfactometer system, and optimized testing conditions for Aedes albopictus (Diptera: Culicidae). We found no significant difference in the biting activity of the insect between 8:00 and 22:00. Furthermore, 5-10 day old mosquitoes were relatively strongly attracted, with bloodsucking rate 75.8%. The random capture rate (entered trap in absence of odor) was less than 20% for the 20-40 tested insects. Capture rates and systematic errors increased as the number of testing insects increased. Wind speed affected the capture rate significantly, whereas variations in temperature between 25-28°C did not result in significant difference. The wind speed of 0.2 m/s exhibited a higher capture rate, which was significantly different from those at either 0.1 m/s or 0.4 m/s (P eugenol, anisaldehyde, geraniol, citronellal, citral, and linalool all exhibited some inhibition effect on mosquitoes to successfully trace human odor or 1% of L-lactic acid in dichloromethane. The results of these two tests indicate that the L-lactic acid/dichloromethane mixture may be used as an effective attractant to evaluate the effect of possible spatial repellents on Ae. albopictus.

Full Text Available Neem (Azadiracta India seed oil in appropriate amount when smeared on the surface of the hand showed excellent repellent action against Aedes aegypti mosquitoes. When 1 ml of oil was spread on the hand, with an approximate area of 160 sq cm the percentage of alighting and blood fed mosquitoes in the experimental cages varied from 14 to 78 and 4 to 46 respectively. This percentage decreased to 6 to 18 and 0 to 16 respectively when the amount of oil applied was 1.5 ml. Only 0-4% of the mosquitoes alighted on the skin of which 2% only took the blood meal when 2 ml of the oil was used to cover the hand. In the control cages cent percent of the mosquitoes alighted and sucked blood. The repellent action was directly proportional to the hour of exposure to the oil. It was also observed that even after alighting on a oil- smeared skin a sizeable proportion of mosquitoes were not able to imbibe blood meal. Neem seed oil was non-toxic, non- irritating to skin.

Full Text Available Objective: To assess the larvicidal potential of the crude benzene, chloroform, ethyl acetate and methanol solvent extracts of the medicinal plant Impatiens balsamina against Anopheles stephensi (An. stephensi, Aedes aegypti (Ae. aegypti and Culex quinquefasciatus (Cx. quinquefasciatus. Methods: Twenty five third instar larvae of An. stephensi, Ae. aegypti and Cx. quinquefasciatus were exposed to various concentrations and were assayed in the laboratory by using the protocol recommended by WHO. The larval mortality was observed after 24 h of treatment. Results: Among extracts tested, the highest larvicidal activity was observed in leaf methanol extract of Impatiens balsamina against An. stephensi, Ae. aegypti and Cx. quinquefasciatus with the LC50 and LC90 values 98.04, 119.68, 125.06 and 172.93, 210.14, 220.60 mg/L, respectively. Conclusions: From the results it can be concluded that the larvicidal effect of Impatiens balsamina against An. stephensi, Ae. aegypti and Cx. quinquefasciatus make this plant product promising as an alternative to synthetic insecticide in mosquito control programs.

Objective: To assess the larvicidal potential of the crude benzene, chloroform, ethyl acetate and methanol solvent extracts of the medicinal plant Impatiens balsamina against Anopheles stephensi(An. stephensi), Aedes aegypti (Ae. aegypti) and Culex quinquefasciatus (Cx. quinquefasciatus). Methods: Twenty five third instar larvae of An. stephensi, Ae. aegypti and Cx. quinquefasciatus were exposed to various concentrations and were assayed in the laboratory by using the protocol recommended by WHO. The larval mortality was observed after 24 h of treatment. Results: Among extracts tested, the highest larvicidal activity was observed in leaf methanol extract of Impatiens balsamina against An. stephensi, Ae. aegypti and Cx. quinquefasciatus with the LC50 and LC90 values 98.04, 119.68, 125.06 and 172.93, 210.14, 220.60 mg/L, respectively. Conclusions:From the results it can be concluded that the larvicidal effect of Impatiens balsamina against An. stephensi, Ae. aegypti and Cx. quinquefasciatus make this plant product promising as an alternative to synthetic insecticide in mosquito control programs.

Full Text Available The Asian tiger mosquito, Aedes albopictus, is a vector of dengue, Chikungunya, and Zika viruses. This mosquito inhabits a wide range of artificial water-holding containers in urban and suburban areas making it difficult to control. We tested the hypothesis that female-driven autodissemination of an insect growth regulator could penetrate cryptic oviposition habitats difficult to treat with conventional insecticidal sprays.Oviposition preferences of Ae. albopictus females for open and cryptic cups were tested in semi-field experiments. Two conventional larvicidal sprayers were tested to determine droplet penetration and larvicidal efficacy in open and cryptic habitats using Bacillus thuringiensis var. israelensis (Bti in the field. Finally, the efficacy of pyriproxyfen autodissemination stations was assessed in cryptic and open cups in residential areas during 2013 and 2014.Gravid females strongly preferred cryptic (53.1±12.9 eggs/cup over open (10.3±4.3 eggs/cup cups for oviposition. Cryptic cups showed limited droplet penetration and produced 0.1-0.3% larval mortality with a conventional backpack and low-volume sprays of Bti. The autodissemination stations effectively contaminated these cryptic cups (59.3-84.6% and produced 29.7-40.8% pupal mortality during 2013-2014. Significant pupal mortality was also observed in open cups.The autodissemination station effectively exploits the oviposition behavior of wild gravid females to deliver pyriproxyfen to targeted oviposition habitats. Although the pupal mortality in cryptic cups was relatively lower than expected for the effective vector control. Autodissemination approach may be a suitable supporting tool to manage Ae. albopictus immatures in the cryptic habitats those are less accessible to conventional larvicidal sprays.

Background The Asian tiger mosquito, Aedes albopictus, is a vector of dengue, Chikungunya, and Zika viruses. This mosquito inhabits a wide range of artificial water-holding containers in urban and suburban areas making it difficult to control. We tested the hypothesis that female-driven autodissemination of an insect growth regulator could penetrate cryptic oviposition habitats difficult to treat with conventional insecticidal sprays. Methodology Oviposition preferences of Ae. albopictus females for open and cryptic cups were tested in semi-field experiments. Two conventional larvicidal sprayers were tested to determine droplet penetration and larvicidal efficacy in open and cryptic habitats using Bacillus thuringiensis var. israelensis (Bti) in the field. Finally, the efficacy of pyriproxyfen autodissemination stations was assessed in cryptic and open cups in residential areas during 2013 and 2014. Principal Findings Gravid females strongly preferred cryptic (53.1±12.9 eggs/cup) over open (10.3±4.3 eggs/cup) cups for oviposition. Cryptic cups showed limited droplet penetration and produced 0.1–0.3% larval mortality with a conventional backpack and low-volume sprays of Bti. The autodissemination stations effectively contaminated these cryptic cups (59.3–84.6%) and produced 29.7–40.8% pupal mortality during 2013–2014. Significant pupal mortality was also observed in open cups. Conclusions The autodissemination station effectively exploits the oviposition behavior of wild gravid females to deliver pyriproxyfen to targeted oviposition habitats. Although the pupal mortality in cryptic cups was relatively lower than expected for the effective vector control. Autodissemination approach may be a suitable supporting tool to manage Ae. albopictus immatures in the cryptic habitats those are less accessible to conventional larvicidal sprays. PMID:28033379

Aedes aegypti mosquito is the principal vector of the viruses responsible for urban yellow fever, dengue, dengue haemorrhagic fever, as well as Zika and chikungunya in Brazil. The present study was aimed to investigate the insecticidal potential of the extract and fractions of Ottonia anisum, along with special metabolites isolated from it, as natural alternatives against larvae (L3) of Ae. aegypti, vector of potentially deadly tropical infections in Brazil. The plant species O. anisum was collected in March 2015, at Xerιm area, in Rio de Janeiro City, Brazil. Crude extracts and the isolated pure compounds were screened for toxicity against Ae. aegypti larvae (L3). Bioassays were performed on 20 larvae (L3) of Ae. aegypti in triplicate. The samples were dissolved in a mixture of acetone and DMSO at final concentrations of 1-200 μg/ml. The toxicity of the solutions was evaluated towards the growth and development of Ae. aegypti larvae till emergence of adults. The crude hexane extract showed 100% larval mortality 24 h after treatment at a concentration of 200 μg/ml. The bioassays using 1-butyl-3,4-methylenedioxybenzene revealed 100% mortality among L3 larvae, 24 h afterthe treatment at a concentration of 30 μg/ml, the LC recorded was 1.6 μg/ml. At concentration of 10 μg/ml, the L3 larval mortality recorded was 92%. The metabolite 1-butyl-3,4-methylenedioxybenzene showed potent toxicity against Ae. aegypti larvae. This arylbutanoid agent could be used as a natural alternative adjuvant pesticide, in new compositions that would be environmentally safer.

Full Text Available BACKGROUND: Control of the mosquito vector population is the most effective strategy currently available for the prevention of dengue fever and the containment of outbreaks. Photo-activated oxidants may represent promising tools for developing effective, safe and ecofriendly novel larvicides. The purpose of this study was to evaluate the potential of the synthetic meso-substituted porphyrin meso-tri(N-methylpyridyl, meso-mono(N-tetradecylpyridylporphine (C14 as a photoactivatable larvicide against the dengue vector Aedes (Stegomyia aegypti. METHODOLOGY: The photophysical and photochemical properties of the C14 molecule were assessed spectrophotometrically. Photomediated larvicidal efficacy, route of intake and site of action were determined on Ae. aegypti larvae by laboratory bioassays and fluorescence microscopy. Using powdered food pellet for laboratory rodents (a common larval food used in the laboratory as a carrier for C14, loading-release dynamics, larvicidal efficacy and residual activity of the C14-carrier complex were investigated. MAIN FINDINGS: The C14 molecule was found to exert a potent photosensitizing activity on Ae. aegypti larvae. At irradiation intervals of 12 h and 1 h, at a light intensity of 4.0 mW/cm(2, which is 50-100 times lower than that of natural sunlight, LC(50 values of 0.1 µM (0.15 mg/l and 0.5 µM (0.77 mg/l were obtained, respectively. The molecule was active after ingestion by the larvae and caused irreversible, lethal damage to the midgut and caecal epithelia. The amphiphilic nature of C14 allowed a formulate to be produced that not only was as active against the larvae as C14 in solution, but also possessed a residual activity of at least two weeks, in laboratory conditions. CONCLUSIONS: The meso-substituted synthetic porphyrin C14, thanks to its photo-sensitizing properties represents an attractive candidate for the development of novel photolarvicides for dengue vector control.

In the summer of 2005, the Asian tiger mosquito, Aedes albopictus (Skuse) (Diptera: Culicidae) was found for the first time in the Netherlands. It was collected on the premises of several horticultural companies that import the ornamental plant Dracaena sanderiana (Sparagalus: Dracaenaceae

In the summer of 2005, the Asian tiger mosquito, Aedes albopictus (Skuse) (Diptera: Culicidae) was found for the first time in the Netherlands. It was collected on the premises of several horticultural companies that import the ornamental plant Dracaena sanderiana (Sparagalus: Dracaenaceae [Agavacea

The Asian tiger mosquitoAedes albopictus (Skuse), is a highly invasive species that continues to expand its geographic distribution both in the United States and in countries on other continents. Studies have demonstrated its susceptibility to infection with at least 32 viruses, including 13 that are present in the United States. Despite this susceptibility, its role as a significant competent vector in natural transmission cycles of arboviruses, has been limited. However, with the recent introductions of chikungunya and Zika viruses into the Americas, for which Ae. albopictus is a recognized vector, it is possible that the species may contribute to the transmission of these viruses to humans and perhaps other susceptible vertebrates.

The generation and study of transgenic Aedes aegypti mosquitoes provides an essential tool for elucidating the complex molecular biology of this important vector. Within the field, genetic manipulation has surpassed the proof of principle stage and is now utilized in both applied and theoretical vector control strategies. The application of new instruments, technologies and techniques allows ever more controlled experiments to be conducted. In this text we describe microinjection of Ae. aegypti embryos in the context of evaluating and performing genomic editing with transcription activator-like effector nucleases (TALENs).

The RNA interference pathway functions as an antiviral defense in invertebrates. In order to generate a phenotypic marker which “senses” the status of the RNAi pathway in Aedes aegypti, transgenic strains were developed to express EGFP and DsRED marker genes in the eye, as well as double-stranded RNA homologous to a portion of the EGFP gene. Transgenic “sensor” mosquitoes exhibited robust eye-specific DsRED expression with little EGFP, indicating RNAi-based silencing. Cloning and high-through...

The generation and study of transgenic Aedes aegypti mosquitoes provides an essential tool for elucidating the complex molecular biology of this important vector. Within the field, genetic manipulation has now surpassed the proof of principle stage and is now utilised in both applied and theoretical vector control strategies. The application of new instruments, technologies and techniques allows ever more controlled experiments to be conducted. In this text we describe microinjection of Ae. aegypti embryos in the context of evaluating and performing genomic editing with transcription activator-like effector nucleases (TALENs). PMID:26443221

Full Text Available Following the epidemic disease caused by the Chikungunya virus detected in the Provinces of Romagna during 2007, a specific monitoring-plan against the bug-vector Aedes albopticus was set up by the Agenzia Regionale Prevenzione e Ambiente dell’Emilia Romagna (ARPA in the he Emilia-Romagna region (Italy. The analytical method consisted in the simple enumeration of the mosquitoes eggs spawned on a appropriate substratum, using an optic microscope.The aim of this study was to guarantee data comparability among the several laboratories involved in the project. Using the Analysis of Variance (ANOVA, homogeneous and comparable analytical data were emphasised.

Mosquito larvicidal activity of Piper longum fruit-derived materials against the fourth-instar larvae of Aedes aegypti was examined. A crude methanol extract of P. longum fruits was found to be active against the larvae, and the hexane fraction of the methanol extract showed a strong larvicidal activity of 100% mortality. The biologically active component of P. longum fruits was characterized as pipernonaline by spectroscopic analyses. The LC(50) value of pipernonaline was 0.25 mg/L. The toxicity of pipernonaline is comparable to that of pirimiphos-methyl as a mosquito larvicide. In tests with available components derived from P. longum, no activity was observed with piperettine, piperine, or piperlongumine.

Extracts from Justicia adhatoda L. (Acanthaceae) strongly reduced the fitness of the mosquito, Aedes aegypti Linn. The methanolic extracts inhibited several enzymes responsible for protecting insects from oxidative and other damage, including glutathione-S-transferase, superoxide dismutase, cytochrome P450, and α- and β-esterases. They increased repellency (maximum repellency at 100 ppm) in host-seeking adult females using the "arm-in cage assay." Histopathological examination showed the extracts led to serious midgut cell damage. Justicia adhatoda extracts led to reduced fecundity and oviposition of gravid females compared to controls. The extracts led to substantially reduced A. aegypti survival. We infer that the extracts have potential to reduce pathogen transmission by suppressing population growth of A. aegypti, and possibly other mosquito species.

In this paper we study the long time dynamics of a reaction diffusion system, describing the spread of Aedes aegypti mosquitoes, which are the primary cause of dengue infection. The system incorporates a control attempt via the sterile insect technique. The model incorporates female mosquitoes sexual preference for wild males over sterile males. We show global existence of strong solution for the system. We then derive uniform estimates to prove the existence of a global attractor in L-2(Omega), for the system. The attractor is shown to be L-infinity(Omega) regular and posess state of extinction, if the injection of sterile males is large enough. We also provide upper bounds on the Hausdorff and fractal dimensions of the attractor.

Full Text Available Objective: To study the diversity and distribution of mosquitoes in Western Ghats of Coimbatore and Nilgiris District, Tamilnadu, India. Methods: Random collections were carried out during August-2013 to July-2014 in cesspits, animal footprints, rock holes, tree holes, drainages at study areas of Marudhamalai, Valparai, Mettupalayam in Coimbatore District and Dhottapeta, Coonoor, Gudalur in Nilgiris District of Tamilnadu, India by using suction tube and kerosene pump. Mosquitoes were identified by standard entomological procedures. Results: A total of 1 018 mosquitoes (larvae and pupae were collected from all over the study areas comprising 6 genera and 23 species. They are, Culex mimulus, Culex pseudovishnui, Culex quinquefasciatus, Culex vishnui, Culex khazani, Culex uniformis, Heizmannia chandi, Heizmannia grenii, Heizmannia indica, Oclerotatus anureostriatus, Oclerotatus albotaeniatis, Oclerotatus deccanus, Oclerotatus gubernatoris, Aedes aegypti, Aedes albopictus, Aedes edwardsi, Aedes krombeini, Toxorhynchites minimus, Toxorhynchites splendens, Anopheles aitkenii, Anopheles barbirostris, Anopheles culiciformis and Anopheles maculatus. ShannonWeaver diversity index, Margalef’s index of richness and Simpsons dominance index was also studied. From 6 sites, the highest mosquitoes were collected from Marudhamalai (309 and the least mosquitoes were collected in Mettupalayam (68. The study determined whether physicochemical characteristics differ between habitats with high and low presence of mosquito larvae. Based on Margalef’s index of richness (Dmg, the highest values were present in Mettupalayam (5.214 study area and the lowest in Marudhamalai (3.837. It can be concluded from Shanon-Weaver index of diversity that, the highest values were present in Mettupalayam (2.947 and the least value were in Gudalur (2.410 during the study period. Conclusions: In areas with reservoirs of disease, mosquito abundance information can help to identify the

not necessary if the downstream goal is to collect, fix and stain hemocytes for diagnostics. Here, we demonstrate our method of collecting mosquito hemolymph that combines the simplicity of perfusion, using anticoagulant buffers in place of saline solution, with the accuracy of high injection techniques to isolate clean preparations of hemocytes in Aedesmosquitoes.

Tango is a transposon of the Tc1 family and was originally discovered in the African malaria mosquito, Anopheles gambiae. Here we report a systematic analysis of the genome sequence of the yellow fever mosquito, Aedes aegypti, which uncovered three distinct Tango transposons. We name the only An. gambiae Tango transposon AgTango1 and the three Ae. aegypti Tango elements AeTango1-3. Like AgTango1, AeTango1 and AeTango2 elements both have members that retain characteristics of autonomous elements such as intact open reading frames and terminal inverted repeats (TIRs). AeTango3 is a degenerate transposon with no full-length members. All full-length Tango transposons contain subterminal direct repeats within their TIRs. AgTango1 and AeTango1-3 form a single clade among other Tc1 transposons. Within this clade, AgTango1 and AeTango1 are closely related and share approximately 80% identity at the amino acid level, which exceeds the level of similarity of the majority of host genes in the two species. A survey of Tango in other mosquito species was carried out using degenerate PCR. Tango was isolated and sequenced in all members of the An. gambiae species complex, Aedes albopictus and Ochlerotatus atropalpus. Oc. atropalpus contains a rich diversity of Tango elements, while Tango elements in Ae. albopictus and the An. gambiae species complex all belong to Tango1. No Tango was detected in Culex pipiens quinquefasciatus, Anopheles stephensi, Anopheles dirus, Anopheles farauti or Anopheles albimanus using degenerate PCR. Bioinformatic searches of the Cx. p. quinquefasciatus (~10 x coverage) and An. stephensi (0.33 x coverage) databases also failed to uncover any Tango elements. Although other evolutionary scenarios cannot be ruled out, there are indications that Tango1 underwent horizontal transfer among divergent mosquito species.

Full Text Available In the last few years tiger mosquitoes (Aedes albopictus, quickly and widely spread in Italy, represent ideal vectors for different Arboviruses, particularly Dengue virus (DenV and Chikungunya virus (ChikV, who are causing millions of patients in the world per year. For ChikV, appeared for the first time in Italy in 2007, a Surveillance Plan was defined in the Marche Region, a neighbour county of the Italian outbreak site. As a support for this surveillance, we decided to create a new multiplex RT-PCR protocol to detect ChikV directly in tiger mosquitoes. All the mosquitoes were collected with BG-Sentinel® traps (Biogents AG, Regensburg, Germany.Total RNA extraction was carried with Helix RNA plus kit (Diatech srl, Jesi, Italy. For retro-transcription and amplification a Mastercycler® ep gradient S thermal cycler (Eppendorf AG, Hamburg, Germany was used. From the whole RNA extracted from captured mosquitoes, we developed a new end-point multiplex retro transcriptase polymerase chain reaction (RT-PCR, for both the detection and identification of Aedes spp. and ChikV. This RT-PCR protocol is able to detect ChikV directly from adult insects, during alerts or emergencies.The entomological trapping associated with bio-molecular methods represents an effective strategy to detect ChikV directly from vectors. Moreover, after a specific evaluation, this RT-PCR protocol could be applied also for human blood samples in regions with the certain presence of this virus.

Although numerous reports are available concerning the larvicidal potential of essential oils, very few investigations have focused on their mechanisms of action. In the present study, we have investigated the chemical composition of the leaf oil of Croton rhamnifolioides during storage and its effects on oviposition and survival of larvae of the dengue fever mosquitoAedes aegypti. In addition, we have established a possible mechanism of action for the larvicidal activity of the essential oil. GC-MS analyses revealed marked differences in the composition of oil that had been freshly isolated and that of a sample that had been stored in a sealed amber-glass vial under refrigeration for three years. However, both fresh and stored oil exhibited substantial larvicidal activities with LC50 values of 122.35 and 89.03 ppm, respectively, and oviposition deterrent effects against gravid females at concentrations of 50 and 100 µg·mL-1. These results demonstrate that the larvicidal effect of the essential oil was unchanged during three years of storage even though its chemical composition altered. Hence, the essential oil could be used in the preparation of commercial products. In addition, we observed that the trypsin-like activity of mosquito larvae was inhibited in vitro by the essential oil of C. rhamnifolioides, suggesting that the larvicidal effect may be associated with inhibition of this enzyme.

The endosymbiotic bacterium Wolbachia pipientis infects many species of insects and has been transinfected into the mosquitoAedes aegypti (L.), the primary vector of dengue virus (DENV). Recently, it has been shown that Wolbachia blocks the replication and transmission of RNA viruses, such as DENV, in a number of mosquito species including Ae. aegypti and Aedes albopictus (Skuse), which is naturally infected with Wolbachia and considered a secondary vector for DENV. The mosquito species Aedes notoscriptus (Skuse) is highly prevalent in Australia, including in areas where DENV outbreaks have been recorded. The mosquito has been implicated in the transmission of Ross River and Barmah Forest viruses, but not DENV. We investigated whether Wolbachia naturally infects this mosquito species and whether it has an impact on the ability of Ae. notoscriptus to transmit DENV. We show, for the first time, that Ae. notoscriptus is naturally infected with a strain of Wolbachia that belongs to supergroup B and is localized only in the ovaries. However, Wolbachia infection in Ae. notoscriptus did not induce resistance to DENV and had no effect on overall DENV infection rate or titer. The presence of a native Wolbachia in Ae. notoscriptus cannot explain why this mosquito is an ineffective vector of DENV.

We examined the relationship between distance from major larval habitats and abundance of adult mosquitoes in the semiarid plains landscape characteristic of eastern Colorado. Mosquito collection was conducted from late June to early August 2007 and included trap locations at distances ranging from Culex tarsalis Coquillett, Ochlerotatus dorsalis (Meigen) (=Ae. dorsalis), Ochlerotatus melanimon (Dyar) (=Ae. melanimon), and Culex pipiens L. Distance to nearest major larval habitat was not strongly related to Culex abundance within the approximately = 400-m range from larval habitats examined in this study. Abundance of Ae. vexans declined significantly with distance from the larval habitat, whereas abundance was significantly higher in the 20-150- and 160-373-m classes compared with areas within 10 m of the larval habitat for both Ochlerotatus species. Except for Ae. vexans, however, we did not find monotonic increasing or decreasing abundance trends associated with distance from larval habitats for the 400-m range examined. This, combined with a finding that fine-scale habitat heterogeneity influenced abundance for most of the mosquitoes examined, underscores the importance of considering not only distance from larval habitat but also fine-scale habitat heterogeneity to understand how important nuisance-biters and West Nile virus (family Flaviviridae, genus Flavivirus, WNV) vectors use the landscape. We also discuss how these results relate to previous studies from western North America and explore their relevance to operational implementation of adulticides to suppress mosquito vectors during WNV disease outbreaks in the Great Plains.

Full Text Available A number of studies have shown that both innate and adaptive immune defense mechanisms greatly influence the course of human dengue virus (DENV infections, but little is known about the innate immune response of the mosquito vector Aedes aegypti to arbovirus infection. We present evidence here that a major component of the mosquito innate immune response, RNA interference (RNAi, is an important modulator of mosquito infections. The RNAi response is triggered by double-stranded RNA (dsRNA, which occurs in the cytoplasm as a result of positive-sense RNA virus infection, leading to production of small interfering RNAs (siRNAs. These siRNAs are instrumental in degradation of viral mRNA with sequence homology to the dsRNA trigger and thereby inhibition of virus replication. We show that although dengue virus type 2 (DENV2 infection of Ae. aegypti cultured cells and oral infection of adult mosquitoes generated dsRNA and production of DENV2-specific siRNAs, virus replication and release of infectious virus persisted, suggesting viral circumvention of RNAi. We also show that DENV2 does not completely evade RNAi, since impairing the pathway by silencing expression of dcr2, r2d2, or ago2, genes encoding important sensor and effector proteins in the RNAi pathway, increased virus replication in the vector and decreased the extrinsic incubation period required for virus transmission. Our findings indicate a major role for RNAi as a determinant of DENV transmission by Ae. aegypti.

MicroRNAs (miRNAs) are endogenous, single-stranded small RNAs that have important regulatory functions at the post-transcriptional level. Here, we characterize miRNAs in two divergent mosquito species, Aedes aegypti and Anopheles stephensi, through deep sequencing of small RNAs spanning all developmental stages. We discovered eight novel miRNAs in Ae. aegypti and 20 novel miRNAs in An. stephensi, which enabled the first systematic analysis of miRNA evolution in mosquitos. We traced the phylogenetic history of all miRNAs in both species and report a rate of 0.055–0.13 miRNA net gain per million years. Most novel miRNAs originate de novo. Duplications that produced miRNA clusters and families are more common in Ae. aegypti than in An. stephensi. We also identified arm-switch as a source of new miRNAs. Expression profile analysis identified mosquito-specific miRNAs that showed strong stage-specific expression in one or both lineages. For example, the aae-miR-2941/2946 family represents the most abundant maternally-deposited and zygotically transcribed miRNAs in Ae. aegypti. miR-2943 is a highly expressed zygotic miRNA in both Ae. aegypti and An. stephensi. Such information provides the basis to study the function of these miRNAs in biology common to all mosquitos or unique to one particular lineage. PMID:25420875

The RNA interference pathway functions as an antiviral defense in invertebrates. In order to generate a phenotypic marker which "senses" the status of the RNAi pathway in Aedes aegypti, transgenic strains were developed to express EGFP and DsRED marker genes in the eye, as well as double-stranded RNA homologous to a portion of the EGFP gene. Transgenic "sensor" mosquitoes exhibited robust eye-specific DsRED expression with little EGFP, indicating RNAi-based silencing. Cloning and high-throughput sequencing of small RNAs confirmed that the inverted-repeat transgene was successfully processed into short-interfering RNAs by the mosquito RNAi pathway. When the A. aegypti homologues of the genes DCR-2 or AGO-2 were knocked down, a clear increase in EGFP fluorescence was observed in the mosquito eyes. Knockdown of DCR-2 was also associated with an increase in EGFP mRNA levels, as determined by Northern blot and real-time PCR. Knockdown of AGO-3, a gene involved in the germline-specific piRNA pathway, did not restore EGFP expression at either the mRNA or protein level. This transgenic sensor strain can now be used to identify other components of the mosquito RNAi pathway and has the potential to be used in the identification of arboviral suppressors of RNAi.

Nepeta essential oil (Neo) (catnip) and its major component, nepetalactone, have long been known to repel insects including mosquitoes. However, the neural mechanisms through which these repellents are detected by mosquitoes, including the yellow fever mosquitoAedes aegypti, an important vector of...

Full Text Available BACKGROUND: The mosquito, Aedes aegypti, is the principal vector of the Dengue and yellow fever viruses. During feeding, an adult female can take up more than its own body weight in vertebrate blood. After a blood meal females excrete large amounts of urine through their excretion system, the Malpighian tubules (MT. Diuresis starts within seconds after the mosquito starts feeding. Aquaporins (AQPs are a family of membrane transporters that regulate the flow of water, glycerol and other small molecules across cellular membranes in both prokaryotic and eukaryotic cells. Our aim was to identify aquaporins that function as water channels, mediating transcellular water transport in MTs of adult female Ae. aegypti. METHODOLOGY/PRINCIPAL FINDINGS: Using a bioinformatics approach we screened genome databases and identified six putative AQPs in the genome of Ae. aegypti. Phylogenetic analysis showed that five of the six Ae. aegypti AQPs have high similarity to classical water-transporting AQPs of vertebrates. Using microarray, reverse transcription and real time PCR analysis we found that all six AQPs are expressed in distinct patterns in mosquito tissues/body parts. AaAQP1, 4, and 5 are strongly expressed in the adult female MT. RNAi-mediated knockdown of the MT-expressed mosquito AQPs resulted in significantly reduced diuresis. CONCLUSIONS/SIGNIFICANCE: Our results support the notion that AQP1, 4, and 5 function as water transporters in the MTs of adult female Ae. aegypti mosquitoes. Our results demonstrate the importance of these AQPs for mosquito diuresis after blood ingestion and highlight their potential as targets for the development of novel vector control strategies.

In larvalAedes aegypti, transcripts of the Rhesus-like glycoproteins AeRh50-1 and AeRh50-2 have been detected in the anal papillae, sites of ammonia (NH3/NH4(+)) excretion; however, these putative ammonia transporters have not been previously localized or functionally characterized. In this study, we show that the AeRh50s co-immunolocalize with apical V-type H(+)-ATPase as well as with basal Na(+)/K(+)-ATPase in the epithelium of anal papillae. The double-stranded RNA-mediated knockdown of AeRh50-1 and AeRh50-2 resulted in a significant reduction in AeRh50 protein abundance in the anal papillae, and this was coupled to decreased ammonia excretion. The knockdown of AeRh50-1 resulted in decreased hemolymph [NH4(+)] and pH whereas knockdown of AeRh50-2 had no effect on these parameters. We conclude that the AeRh50s are important contributors to ammonia excretion at the anal papillae of larval A. aegypti, which may be the basis for their ability to inhabit areas with high ammonia levels.

Classifying and quantifying mosquito activity includes a plethora of categories, ranging from measuring flight speeds, repellency, feeding rates, and specific behaviors such as home entry, swooping and resting, among others. Entomologists have been progressing more toward using machine vision for efficiency for this endeavor. Digital methods have been used to study the behavior of insects in labs, for instance via three-dimensional tracking with specialized cameras to observe the reaction of mosquitoes towards human odor, heat and CO2, although virtually none was reported for several important fields, such as repellency studies which have a significant need for a proper response quantification. However, tracking mosquitoes individually is a challenge and only limited number of specimens can be studied. Although tracking large numbers of individual insects is hailed as one of the characteristics of an ideal automated image-based tracking system especially in 3D, it also is a costly method, often requiring specialized hardware and limited access to the algorithms used for mapping the specimens. The method proposed contributes towards (a) unlimited open source use, (b) a low-cost setup, (c) complete guide for any entomologist to adapt in terms of hardware and software, (d) simple to use, and (e) a lightweight data output for collective behavior analysis of mosquitoes. The setup is demonstrated by testing a simple response of mosquitoes in the presence of human odor versus control, one session with continuous human presence as a stimuli and the other with periodic presence. A group of female Aedes aegypti (Linnaeus) mosquitoes are released into a white-background chamber with a transparent acrylic panel on one side. The video feed of the mosquitoes are processed using filtered contours in a threshold-adjustable video. The mosquitoes in the chamber are mapped on the raster where the coordinates of each mosquito are recorded with the corresponding timestamp. The average

Mosquitoes transmit dreadful diseases to human beings wherein biological control of these vectors using plant-derived molecules would be an alternative to reduce mosquito population. Aqueous leaf extract and green synthesized silver nanoparticles (Ag NPs) from Pedilanthus tithymaloides (L.) Poit. were investigated for their efficacy against the dengue vector Aedes aegypti L. (Diptera; Culicidae). The biologically synthesized Ag NPs were characterized by UV-vis spectrum, X-ray diffraction, Fourier transform infrared, and surface characteristics by atomic force microscopy. Further, on exposure of the larvae to varying concentrations of aqueous leaf and Ag NPs for 24 h, these Ag NPs showed 100 % mortality from first to fourth instars and pupae of A. aegypti at 0.25 %, which is the highest concentration, tested, wherein it was the lowest concentration of aqueous leaf extract alone which showed only 10-18 % of mortality. Lethal concentration (LC(50)) values of Ag NPs against the larval and pupal stages were 0.029, 0.027, 0.047, 0.086, and 0.018 % with no mortality in control. These results suggest that the use of P. tithymaloides silver nanoparticles can be a rapid, environmentally safer bio-pesticide which can form a novel approach to develop effective biocides for controlling the target vector.

During an entomological investigation carried out in Bucharest and surroundings in fall of 2012, 45 adult mosquitoes (38 females and 7 males) of Aedes albopictus were collected in a neighborhood from the southern area of the city. The morphological identification of the species was further confirmed by sequencing 2 mitochondrial DNA markers: the cytochrome c oxidase subunit I and NADH dehydrogenase subunit 5 genes. Aedes albopictus was collected again in 2013 in the same area from July until October. During late summer the species was found also in another location in the city, downtown Bucharest. Larvae were found in water barrels and other types of household containers, as well as in rain catch basins. In 2014, following a nuisance complaint of a Bucharest inhabitant, the entomological investigation found aggressive Ae. albopictus adults on his property that harbored many mosquito larvae in container-type breeding habitats. These findings are the 1st records of this invasive species and of its breeding population in Romania, and show maintenance of the species over 2 winter seasons. Surveillance of the species outside the area of the capital city was not performed, therefore it is not known whether Ae. albopictus has been introduced in other regions of the country. The presence of Ae. albopictus has been reported every year (2012-14) to competent public health authorities, stressing on the importance of surveillance and of implementation of control measures.

Dengue fever (DF) is the most prevalent arthropod-borne viral disease which affects humans. DF is caused by the four dengue virus (DENV) serotypes, which are transmitted to the host by the mosquitoAedes aegypti that has key roles in DENV infection, replication, and viral transmission (vector competence). Mosquito saliva also plays an important role during DENV transmission. In this study, we detected the presence of sialic acid (Sia) in Aedes aegypti tissues, which may have an important role during DENV-vector competence. We also identified genome sequences encoding enzymes involved in Sia pathways. The cDNA for Aedes aegypti CMP-Sia synthase (CSAS) was amplified, cloned, and functionally evaluated via the complementation of LEC29.Lec32 CSAS-deficient CHO cells. AedesCSAS-transfected LEC29.Lec32 cells were able to express Sia moieties on the cell surface. Sequences related to α-2,6-sialyltransferase were detected in the Aedes aegypti genome. Likewise, we identified Sia-α-2,6-DENV interactions in different mosquito tissues. In addition, we evaluated the possible role of sialylated molecules in a salivary gland extract during DENV internalization in mammalian cells. The knowledge of early DENV-host interactions could facilitate a better understanding of viral tropism and pathogenesis to allow the development of new strategies for controlling DENV transmission.

Dengue fever (DF) is the most prevalent arthropod-borne viral disease which affects humans. DF is caused by the four dengue virus (DENV) serotypes, which are transmitted to the host by the mosquitoAedes aegypti that has key roles in DENV infection, replication, and viral transmission (vector competence). Mosquito saliva also plays an important role during DENV transmission. In this study, we detected the presence of sialic acid (Sia) in Aedes aegypti tissues, which may have an important role during DENV-vector competence. We also identified genome sequences encoding enzymes involved in Sia pathways. The cDNA for Aedes aegypti CMP-Sia synthase (CSAS) was amplified, cloned, and functionally evaluated via the complementation of LEC29.Lec32 CSAS-deficient CHO cells. AedesCSAS-transfected LEC29.Lec32 cells were able to express Sia moieties on the cell surface. Sequences related to α-2,6-sialyltransferase were detected in the Aedes aegypti genome. Likewise, we identified Sia-α-2,6-DENV interactions in different mosquito tissues. In addition, we evaluated the possible role of sialylated molecules in a salivary gland extract during DENV internalization in mammalian cells. The knowledge of early DENV-host interactions could facilitate a better understanding of viral tropism and pathogenesis to allow the development of new strategies for controlling DENV transmission. PMID:25874215

Full Text Available Dengue fever (DF is the most prevalent arthropod-borne viral disease which affects humans. DF is caused by the four dengue virus (DENV serotypes, which are transmitted to the host by the mosquitoAedes aegypti that has key roles in DENV infection, replication, and viral transmission (vector competence. Mosquito saliva also plays an important role during DENV transmission. In this study, we detected the presence of sialic acid (Sia in Aedes aegypti tissues, which may have an important role during DENV-vector competence. We also identified genome sequences encoding enzymes involved in Sia pathways. The cDNA for Aedes aegypti CMP-Sia synthase (CSAS was amplified, cloned, and functionally evaluated via the complementation of LEC29.Lec32 CSAS-deficient CHO cells. AedesCSAS-transfected LEC29.Lec32 cells were able to express Sia moieties on the cell surface. Sequences related to α-2,6-sialyltransferase were detected in the Aedes aegypti genome. Likewise, we identified Sia-α-2,6-DENV interactions in different mosquito tissues. In addition, we evaluated the possible role of sialylated molecules in a salivary gland extract during DENV internalization in mammalian cells. The knowledge of early DENV-host interactions could facilitate a better understanding of viral tropism and pathogenesis to allow the development of new strategies for controlling DENV transmission.

The mosquito species Aedes aegypti is the primary vector of many arboviral diseases, including dengue and yellow fevers, that are responsible for a large worldwide health burden. The biological rhythms of mosquitoes regulate many of the physiological processes and behaviors that influence the transmission of these diseases. For insight into the molecular basis of biological rhythms, diel and circadian gene expression profiling has been carried out for many species. To bring these resources to Aedes aegypti researchers, we used microarray technology to carry out a genome wide assessment of gene expression during the 24 hour light/dark (LD) cycle and during constant darkness (DD). The purpose of this report is to describe the methods, the validation of the results, and the organization of this database resource. The Aedes aegypti Circadian Database is a publicly accessible database that can be searched via a text-based query to visualize 44 hour temporal expression patterns of a given gene in Ae. aegypti heads under diel (observed under a 12 hour/12 hour LD cycle) and circadian (observed under DD) conditions. Profiles of gene expression under these conditions were assayed by Nimblegen 12-plex microarrays and rhythmicity was objectively assessed by the JTK_CYCLE algorithm. The output of the search is a graphical representation of the expression data along with computed period length, the time-of-day of gene expression peaks, and statistical determination for rhythmicity. Our results show that at least 7.9% of the gene set present in the Aedes aegypti head are rhythmic under LD conditions and 6.7% can be considered circadian, oscillating under constant dark conditions. We present these results in the Aedes aegypti Circadian Database through Bioclock, a public website hosted by the University of Notre Dame at http://www.nd.edu/~bioclock/. This website allows searchable browsing of this quantitative gene expression information. The visualization allows for gene

Open marsh water management (OMWM) is a commonly used approach to manage salt marsh mosquitoes than can obviate the need for pesticide application and at the same time, partially restore natural functions of grid-ditched marshes. OMWM includes a variety of hydrologic manipulations, often tailored to the specific conditions on individual marshes, so the overall effectiveness of this approach is difficult to assess. Here, we report the results of controlled field trials to assess the effects of two approaches to OMWM on larvalmosquito production at National Wildlife Refuges (NWR). A traditional OMWM approach, using pond construction and radial ditches was used at Edwin B. Forsythe NWR in New Jersey, and a ditch-plugging approach was used at Parker River NWR in Massachusetts. Mosquito larvae were sampled from randomly placed stations on paired treatment and control marshes at each refuge. The proportion of sampling stations that were wet declined after OMWM at the Forsythe site, but not at the Parker River site. The proportion of samples with larvae present and mean larval densities, declined significantly at the treatment sites on both refuges relative to the control marshes. Percentage of control for the 2 yr posttreatment, compared with the 2 yr pretreatment, was >90% at both treatment sites.

Background: Chikungunya virus (CHIKV) is an arthritogenic alphavirus (family Togaviridae), transmitted by Aedes species mosquitoes. CHIKV re-emerged in 2004 with multiple outbreaks worldwide and recently reached the Americas where it has infected over a million individuals in a rapidly expanding

BACKGROUND: Chikungunya virus (CHIKV) is an arthritogenic alphavirus (family Togaviridae), transmitted by Aedes species mosquitoes. CHIKV re-emerged in 2004 with multiple outbreaks worldwide and recently reached the Americas where it has infected over a million individuals in a rapidly expanding epi

Three gustatory receptor neurons were characterized for contact chemoreceptive sensilla on the labella of female yellow fever mosquitoes, Aedes aegypti. The neuron with the smallest amplitude spike responded to the feeding deterrent, quinine, as well as DEET and other insect repellents. Two other ...

BACKGROUND: Chikungunya virus (CHIKV) is an arthritogenic alphavirus (family Togaviridae), transmitted by Aedes species mosquitoes. CHIKV re-emerged in 2004 with multiple outbreaks worldwide and recently reached the Americas where it has infected over a million individuals in a rapidly expanding epi

Background: Chikungunya virus (CHIKV) is an arthritogenic alphavirus (family Togaviridae), transmitted by Aedes species mosquitoes. CHIKV re-emerged in 2004 with multiple outbreaks worldwide and recently reached the Americas where it has infected over a million individuals in a rapidly expanding

The Asian tiger mosquito, Aedes albopictus, is among the most invasive species in the world. Established in the U.S. since 1985, this species now infests 30 states and continues to spread internationally. Concerned public health officials recognize this species as an important vector of chikunguny...

The Asian tiger mosquito, Aedes albopictus, became established in the continental US in 1985 and now infests 30 states. In 2007 the USDA Agricultural Research Service funded an “area-wide” project focused on the management of this species. The project was a unique federal, state, local collaborati...

Insect kinins (leucokinins) are multifunctional peptides acting as neurohormones and neurotransmitters. In females of the mosquito vector Aedes aegypti (L.), aedeskinins are known to stimulate fluid secretion from the renal organs (Malpighian tubules) and hindgut contractions by activating a G prot...

The Asian tiger mosquito, Aedes albopictus, is a highly successful invasive species that transmits a number of human viral diseases, including dengue and Chikungunya fevers. This species has a large genome with significant population-based size variation. The complete genome sequence was determined

Container-inhabiting Aedes are among the most medically important mosquito vectors of diseases. They also impact health and quality of life by their persistent and severe biting. Monitoring of container-inhabiting Aedes species is challenging due to the need for specialized traps and lures. Biogents Sentinel (BGS) trap has become a standard for Aedes albopictus (Skuse) surveillance; however, it has substantial problems with durability, quality of construction, and sample exposure to the elements. The goal of this study was to develop a methodology for collecting medically important container-inhabiting Aedes species in numbers sufficient for population trend analysis, control efficacy studies, and pathogen testing. Mosquito Magnets (MM) baited with BG lure and R-octenol were selected as the most practical alternative to BGS, collecting significantly more Ae. albopictus (32.1 ± 0.7 vs. 5.6 ± 0.1), Aedes japonicus (Theobald) (10.1 ± 0.4 vs. 1.2 ± 0.02), and Aedes triseriatus (Say) (0.9 ± 0.04 vs. 0.04 ± 0.004) females on average per trapping under a variety of weather conditions. MM can be particularly useful for long-term surveillance or when large numbers of specimens are required for pathogen isolation, such as at the sites with suspected dengue or chikungunya transmission.

Background Viral diseases transmitted via Aedesmosquitoes are on the rise, such as Zika, dengue, and chikungunya. Novel tools to mitigate Aedesmosquitoes-transmitted diseases are urgently needed. We tested whether commercially insecticide-impregnated school uniforms can reduce dengue incidence in school children. Methods We designed a cluster-randomised controlled trial in Thailand. The primary endpoint was laboratory-confirmed dengue infections. Secondary endpoints were school absenteeism; and impregnated uniforms’ 1-hour knock-down and 24 hour mosquito mortality as measured by standardised WHOPES bioassay cone tests at baseline and after repeated washing. Furthermore, entomological assessments inside classrooms and in outside areas of schools were conducted. Results We enrolled 1,811 pupils aged 6–17 from 5 intervention and 5 control schools. Paired serum samples were obtained from 1,655 pupils. In the control schools, 24/641 (3.7%) and in the intervention schools 33/1,014 (3.3%) students had evidence of new dengue infections during one school term (5 months). There was no significant difference in proportions of students having incident dengue infections between the intervention and control schools, with adjustment for clustering by school. WHOPES cone tests showed a 100% knock down and mortality of Aedes aegypti mosquitoes exposed to impregnated clothing at baseline and up to 4 washes, but this efficacy rapidly declined to below 20% after 20 washes, corresponding to a weekly reduction in knock-down and mosquito mortality by 4.7% and 4.4% respectively. Results of the entomological assessments showed that the mean number of Aedes aegypti mosquitoes caught inside the classrooms of the intervention schools was significantly reduced in the month following the introduction of the impregnated uniforms, compared to those collected in classrooms of the control schools (p = 0.04) Conclusions Entomological assessments showed that the intervention had some impact on

Full Text Available Viral diseases transmitted via Aedesmosquitoes are on the rise, such as Zika, dengue, and chikungunya. Novel tools to mitigate Aedesmosquitoes-transmitted diseases are urgently needed. We tested whether commercially insecticide-impregnated school uniforms can reduce dengue incidence in school children.We designed a cluster-randomised controlled trial in Thailand. The primary endpoint was laboratory-confirmed dengue infections. Secondary endpoints were school absenteeism; and impregnated uniforms' 1-hour knock-down and 24 hour mosquito mortality as measured by standardised WHOPES bioassay cone tests at baseline and after repeated washing. Furthermore, entomological assessments inside classrooms and in outside areas of schools were conducted.We enrolled 1,811 pupils aged 6-17 from 5 intervention and 5 control schools. Paired serum samples were obtained from 1,655 pupils. In the control schools, 24/641 (3.7% and in the intervention schools 33/1,014 (3.3% students had evidence of new dengue infections during one school term (5 months. There was no significant difference in proportions of students having incident dengue infections between the intervention and control schools, with adjustment for clustering by school. WHOPES cone tests showed a 100% knock down and mortality of Aedes aegypti mosquitoes exposed to impregnated clothing at baseline and up to 4 washes, but this efficacy rapidly declined to below 20% after 20 washes, corresponding to a weekly reduction in knock-down and mosquito mortality by 4.7% and 4.4% respectively. Results of the entomological assessments showed that the mean number of Aedes aegypti mosquitoes caught inside the classrooms of the intervention schools was significantly reduced in the month following the introduction of the impregnated uniforms, compared to those collected in classrooms of the control schools (p = 0.04.Entomological assessments showed that the intervention had some impact on the number of Aedes

Global warming has a marked influence on the life cycle of epidemic vectors as well as their interactions with human beings. The Aedes albopictus mosquito as the vector of dengue fever surged exponentially in the last decade, raising ecological and epistemological concerns of how climate change altered its growth rate and population dynamics. As the global warming pattern is considerably uneven across four seasons, with a confirmed stronger effect in winter, an emerging need arises as to exploring how the seasonal warming effects influence the annual development of Ae. albopictus. The model consolidates a 35-year climate dataset and designs fifteen warming patterns that increase the temperature of selected seasons. Based on a recently developed mechanistic population model of Ae. albopictus, the model simulates the thermal reaction of blood-fed adults by systematically increasing the temperature from 0.5 to 5 °C at an interval of 0.5 °C in each warming pattern. The results show the warming effects are different across seasons. The warming effects in spring and winter facilitate the development of the species by shortening the diapause period. The warming effect in summer is primarily negative by inhibiting mosquito development. The warming effect in autumn is considerably mixed. However, these warming effects cannot carry over to the following year, possibly due to the fact that under the extreme weather in winter the mosquito fully ceases from development and survives in terms of diapause eggs. As the historical pattern of global warming manifests seasonal fluctuations, this study provides corroborating and previously ignored evidence of how such seasonality affects the mosquito development. Understanding this short-term temperature-driven mechanism as one chain of the transmission events is critical to refining the thermal reaction norms of the epidemic vector under global warming as well as developing effective mosquito prevention and control strategies.

To be transmitted by its mosquito vector, dengue virus (DENV) must infect midgut epithelial cells, replicate and disseminate into the hemocoel, and finally infect the salivary glands, which is essential for transmission. The extrinsic incubation period (EIP) is very relevant epidemiologically and is the time required from the ingestion of virus until it can be transmitted to the next vertebrate host. The EIP is conditioned by the kinetics and tropisms of virus replication in its vector. Here we document the virogenesis of DENV-2 in newly-colonized Aedes aegypti mosquitoes from Chetumal, Mexico in order to understand better the effect of vector-virus interactions on dengue transmission. After ingestion of DENV-2, midgut infections in Chetumal mosquitoes were characterized by a peak in virus titers between 7 and 10 days post-infection (dpi). The amount of viral antigen and viral titers in the midgut then declined, but viral RNA levels remained stable. The presence of DENV-2 antigen in the trachea was positively correlated with virus dissemination from the midgut. DENV-2 antigen was found in salivary gland tissue in more than a third of mosquitoes at 4 dpi. Unlike in the midgut, the amount of viral antigen (as well as the percent of infected salivary glands) increased with time. DENV-2 antigen also accumulated and increased in neural tissue throughout the EIP. DENV-2 antigen was detected in multiple tissues of the vector, but unlike some other arboviruses, was not detected in muscle. Our results suggest that the EIP of DENV-2 in its vector may be shorter that the previously reported and that the tracheal system may facilitate DENV-2 dissemination from the midgut. Mosquito organs (e.g. midgut, neural tissue, and salivary glands) differed in their response to DENV-2 infection.

Full Text Available Abstract Background To be transmitted by its mosquito vector, dengue virus (DENV must infect midgut epithelial cells, replicate and disseminate into the hemocoel, and finally infect the salivary glands, which is essential for transmission. The extrinsic incubation period (EIP is very relevant epidemiologically and is the time required from the ingestion of virus until it can be transmitted to the next vertebrate host. The EIP is conditioned by the kinetics and tropisms of virus replication in its vector. Here we document the virogenesis of DENV-2 in newly-colonized Aedes aegypti mosquitoes from Chetumal, Mexico in order to understand better the effect of vector-virus interactions on dengue transmission. Results After ingestion of DENV-2, midgut infections in Chetumal mosquitoes were characterized by a peak in virus titers between 7 and 10 days post-infection (dpi. The amount of viral antigen and viral titers in the midgut then declined, but viral RNA levels remained stable. The presence of DENV-2 antigen in the trachea was positively correlated with virus dissemination from the midgut. DENV-2 antigen was found in salivary gland tissue in more than a third of mosquitoes at 4 dpi. Unlike in the midgut, the amount of viral antigen (as well as the percent of infected salivary glands increased with time. DENV-2 antigen also accumulated and increased in neural tissue throughout the EIP. DENV-2 antigen was detected in multiple tissues of the vector, but unlike some other arboviruses, was not detected in muscle. Conclusion Our results suggest that the EIP of DENV-2 in its vector may be shorter that the previously reported and that the tracheal system may facilitate DENV-2 dissemination from the midgut. Mosquito organs (e.g. midgut, neural tissue, and salivary glands differed in their response to DENV-2 infection.

Full Text Available Introduction The present study shows the colonization of Aedesmosquitoes in breeding sites specific for Culex quinquefasciatus in neighborhoods in the municipality of Olinda. Methods Samples were collected between May 2011 and June 2012 from breeding sites positive for Cx. quinquefasciatus by using a ladle and manual suction pump. Results Aedes aegypti (0.12%, Aedes albopictus (0.03%, and Cx. quinquefasciatus (99.8% were found across the breeding sites. Conclusions The presence of Aedes ssp. in several Cx. quinquefasciatus breeding sites with a heavy load of organic material demonstrates the need to review the concepts and methods used for treatment, as the use of specific larvicide for breeding sites of Culex.

Chikungunya fever is an emerging mosquito-borne disease caused by the infection with chikungunya virus (CHIKV). The CHIKV has been rarely detected in mosquito vectors from Northern India, since vector surveillance is an effective strategy in controlling and preventing CHIKV transmission. Thus, virological investigation for CHIKV among mosquitoes of Aedes (A.) species was carried out in the Lucknow district during March 2010 to October 2011. We collected adult mosquitoes from areas with CHIKV positive patients. The adult Aedesmosquito samples were pooled, homogenized, clarified and tested for CHIKV by nonstructural protein 1 (nsP1) gene based polymerase chain reaction (PCR). A total 91 mosquito pools comprising of adult A. aegypti and A. albopictus were tested for CHIKV. The partial envelope protein (E1) gene sequences of mosquito-borne CHIKV strains were analyzed for genotyping. Of 91 pools, 6 pools of A. aegypti; and 2 pools of A. albopictus mosquitoes were identified positive for CHIKV by PCR. The phylogenetic analysis revealed clustering of CHIKV strains in two sub-lineages within the monophyletic East-Central South African (ECSA) genotype. Novel amino acid changes at the positions 294 (P294L) and 295 (S295F) were observed during analysis of amino acid sequence of the partial E1 gene. This study demonstrates the genetic diversity of circulating CHIKV strains and reports the first detection of CHIKV strains in Aedes vector species from the state of Uttar Pradesh. These findings have implication for vector control strategies to mitigate vector population to prevent the likelihood of CHIKV epidemic in the near future.

Bamboo stumps can be a congenial breeding habitat of the mosquitoes.In view of this,a preliminary assessment of the dipteran immatures inhabiting the stumps of bamboo groves in the Darjeeling Himalayas was carried out at a spatial scale.Of the 104 stumps of Dendrocalamus hamiltoni surveyed,70 were found to host immatures of three dipteran species,the mosquitoesAedes aegypti and Culex quinquefasciatus and the midges Chironomus sp.in varying densities.Though the stumps varied in diameter,in each stump on average 12.1 immatures were found.The abundance of the immatures was positively correlated with the diameter of the stumps (r = +0.382;P ＜ 0.001) but negatively with the pH of the water present in the stumps (r = -0.336;P ＜ 0.01).The coefficient of association was found to be +8.4 for the Ae.aegypti and Chironomus immatures,while in the rest of the species pair the association seemed to be independent.Thus it can be concluded that the stumps in the bamboo groves of Darjeeling Himalayas provides a favourable habitat for the mosquito and chironomid immatures.

Full Text Available Abstract Background As the ecology of mosquito larvae can be complex there is need to develop a rational framework for undertaking larval ecological studies. Local environmental characteristics, such as altitude, climate and land use, can significantly impact on phenology and population dynamics of mosquito larvae, and indirectly affect the dynamics of mosquito-borne diseases. The aim of this study was to assess the feasibility of implementing an integrated approach to larval source management under the distinct ecological settings. Methods The study was conducted in two highland villages and one village, at a lower altitude, in the Lake Victoria basin, where malaria is endemic and transmitted by the same Anopheles mosquito species. In each village the stability of mosquitolarval habitats was classified as either temporary or permanent. The productivity of these habitat types was quantified by carrying out weekly larval sampling using a standard dipping method for a period of two years. During sampling the physical characteristic of the larval habitat, including the vegetation cover were noted. Ambient temperature, rainfall and relative humidity were recorded on a 21 × Micro-datalogger in each study site. Results Anopheles gambiae sensu lato larvae were found in all study sites. Anopheles arabiensis was more abundant (93% in Nyalenda (Lake Victoria basin and Fort Ternan (highland area; 71%. In Lunyerere (highland area, An. gambiae sensu stricto comprised 93% of the total An. gambiae s.l. larvae. Larvae of An. gambiae s.l. mosquitoes were present in both temporary and permanent habitats with monthly variations dependent on rainfall intensity and location. Anopheles larvae were more likely to be found in man-made as opposed to natural habitats. Grassy habitats were preferred and were, therefore, more productive of Anopheles larvae compared to other habitat types. Weekly rainfall intensity led to an increase or decrease in mosquitolarval

Full Text Available The way in which vectors distribute themselves amongst their hosts has important epidemiological consequences. While the role played by active host choice is largely unquestioned, current knowledge relates mostly to the innate response of vectors towards stimuli signalling the presence or quality of their hosts. Many of those cues, however, can be unpredictable, and therefore prevent the incorporation of the appropriate response into the vector's behavioural repertoire unless some sort of associative learning is possible. We performed a wide range of laboratory experiments to test the learning abilities of the mosquito, Aedes aegypti. Mosquitoes were exposed to choice procedures in (1 an olfactomenter and (2 a 'visual arena'. Our goal was to determine whether the mosquitoes were able to associate unconditional stimuli (blood feeding, human breath, vibration and electrical shock with particular odours (citral, carvone, citronella oil and eugenol and visual patterns (horizontal or vertical black bars to which they had been previously observed to be responsive. We found no evidence supporting the hypothesis that associative learning abilities are present in adult Ae. aegypti. We discuss the possibilities that the assays employed were either inappropriate or insufficient to detect associative learning, or that associative learning is not possible in this species.

Hemocytes are an essential component of the mosquito immune system but current knowledge of the types of hemocytes mosquitoes produce, their relative abundance, and their functions is limited. Addressing these issues requires improved methods for collecting and maintaining mosquito hemocytes in vitro, and comparative data that address whether important vector species produce similar or different hemocyte types. Toward this end, we conducted a comparative study with Anopheles gambiae and Aedes aegypti. Collection method greatly affected the number of hemocytes and contaminants obtained from adult females of each species. Using a collection method called high injection/recovery, we concluded that hemolymph from An. gambiae and Ae. aegypti adult females contains three hemocyte types (granulocytes, oenocytoids and prohemocytes) that were distinguished from one another by a combination of morphological and functional markers. Significantly more hemocytes were recovered from An. gambiae females than Ae. aegypti. However, granulocytes were the most abundant cell type in both species while oenocytoids and prohemocytes comprised less than 10% of the total hemocyte population. The same hemocyte types were collected from larvae, pupae and adult males albeit the absolute number and proportion of each hemocyte type differed from adult females. The number of hemocytes recovered from sugar fed females declined with age but blood feeding transiently increased hemocyte abundance. Two antibodies tested as potential hemocyte markers (anti-PP06 and anti-Dox-A2) also exhibited alterations in staining patterns following immune challenge with the bacterium Escherichia coli.

Males of Aedes aegypti mosquitoes formed swarms in the laboratory, triggered by the onset of the photophase or by the presence of odors from a rat (which is a potential host for females). The swarm attracted both males and females and increased mating activity. The number of copulas per mosquito was positively correlated with the number of mosquitoes in the swarm and with the duration of the swarm. Swarming and mating activity increased with the presence of a host for females. Young sexually immature males, less than 24 h old, flew but did not swarm nor copulate. Observations using an olfactometer showed that swarming males produced a volatile pheromone that stimulates the flying activity of females at a distance. Females also produce a volatile attractant. The results suggest that males, and possibly also females, produce an aggregation pheromone that attracts males and females towards the swarm. The characteristics of the pheromone-mediated swarm may be described as a 3-dimensional lek. Our results suggest that the development of pheromone-based control systems and/or pheromone traps for the monitoring of vector populations is feasible, adding a new tool to combat this vector of several human pathogens.

Mosquito (Aedes togoi) larva has to float its siphon on the water surface to breathe air. To elucidate the floating mechanism of the siphon, morphological structures, especially the flap-like lobes and spiracle of the siphon, were observed. Wettability and dynamic behavior of the lobes on the water surface were also experimentally examined. The lobes formed a hollow cone shape under water and expanded on the water surface. The spiracle was located at the base of the cone. The lobes exhibited hydrophobic wettability. During floating process, the lobes were spread into a triangular shape in apical view, and the spiracle was exposed to air. When they were submerged in water, the lobes were folded into a cone shape to seal the spiracle and protect it from water penetration. These dynamic processes occurred at the water surface. In this study, the floating mechanism of the lobes for respiration was described based on the cross-sectional morphology of the lobes with the pressure induced by surface tension and the hydrostatic pressure proportional to depth. This study would be helpful in elucidating the behavior of mosquito larvae near the water surface and may facilitate the formulation of methods to control mosquito larvae.

New mosquito control strategies centred on the modifying of populations require knowledge of existing population densities at release sites and an understanding of breeding site ecology. Using a quantitative pupal survey method, we investigated production of the dengue vector Aedes aegypti (L.) (Stegomyia aegypti) (Diptera: Culicidae) in Cairns, Queensland, Australia, and found that garden accoutrements represented the most common container type. Deliberately placed 'sentinel' containers were set at seven houses and sampled for pupae over 10 weeks during the wet season. Pupal production was approximately constant; tyres and buckets represented the most productive container types. Sentinel tyres produced the largest female mosquitoes, but were relatively rare in the field survey. We then used field-collected data to make estimates of per premises population density using three different approaches. Estimates of female Ae. aegypti abundance per premises made using the container-inhabiting mosquito simulation (CIMSiM) model [95% confidence interval (CI) 18.5-29.1 females] concorded reasonably well with estimates obtained using a standing crop calculation based on pupal collections (95% CI 8.8-22.5) and using BG-Sentinel traps and a sampling rate correction factor (95% CI 6.2-35.2). By first describing local Ae. aegypti productivity, we were able to compare three separate population density estimates which provided similar results. We anticipate that this will provide researchers and health officials with several tools with which to make estimates of population densities.

Eleven microsatellite markers were used to determine the genetic population structure and spread of Aedes aegypti (Stegomyia aegypti) (Diptera: Culicidae) in Pakistan using mosquitoes collected from 13 different cities. There is a single genetic cluster of Ae. aegypti in Pakistan with a pattern of isolation by distance within the population. The low level of isolation by distance suggests the long-range passive dispersal of this mosquito, which may be facilitated by the tyre trade in Pakistan. A decrease in genetic diversity from south to north suggests a recent spread of this mosquito from Karachi. A strong negative correlation between genetic distance and the quality of road connections shows that populations in cities connected by better road networks are less differentiated, which suggests the human-aided passive dispersal of Ae. aegypti in Pakistan. Dispersal on a large spatial scale may facilitate the strategy of introducing transgenic Ae. aegypti or intracellular bacteria such as Wolbachia to control the spread of dengue disease in Pakistan, but it also emphasizes the need for simple measures to control container breeding sites.

The way in which vectors distribute themselves amongst their hosts has important epidemiological consequences. While the role played by active host choice is largely unquestioned, current knowledge relates mostly to the innate response of vectors towards stimuli signalling the presence or quality of their hosts. Many of those cues, however, can be unpredictable, and therefore prevent the incorporation of the appropriate response into the vector's behavioural repertoire unless some sort of associative learning is possible. We performed a wide range of laboratory experiments to test the learning abilities of the mosquito, Aedes aegypti. Mosquitoes were exposed to choice procedures in (1) an olfactomenter and (2) a 'visual arena'. Our goal was to determine whether the mosquitoes were able to associate unconditional stimuli (blood feeding, human breath, vibration and electrical shock) with particular odours (citral, carvone, citronella oil and eugenol) and visual patterns (horizontal or vertical black bars) to which they had been previously observed to be responsive. We found no evidence supporting the hypothesis that associative learning abilities are present in adult Ae. aegypti. We discuss the possibilities that the assays employed were either inappropriate or insufficient to detect associative learning, or that associative learning is not possible in this species.

West Nile virus (WNV) is a zoonotic arboviral pathogen transmitted by mosquitoes in a cycle that involves wild birds as reservoir hosts. The virus is responsible for outbreaks of viral encephalitis in humans and horses. In Europe, Culex pipiens (Diptera: Culicidae) is considered to be the main vector of WNV, but other species such as Stegomyia albopicta (=Aedes albopictus) (Diptera: Culicidae) may also act as competent vectors of this virus. Since 2008 human cases of WNV disease have been reported in northeast Italy. In 2011, new areas of southern Italy became involved and a first outbreak of WNV lineage 1 occurred on the island of Sardinia. On the assumption that a potential involvement of St. albopicta in WNV transmission cannot be excluded, and in order to evaluate the competence of this species for the virus, an experimental infection of an St. albopicta laboratory colony, established from mosquitoes collected in Sardinia, was carried out. The results were compared with those obtained in a colony of the main vector Cx. pipiens. The study showed St. albopicta collected on Sardinia to be susceptible to WNV infection, which suggests this Italian mosquito species is able to act as a possible secondary vector, particularly in urban areas where the species reaches high levels of seasonal abundance.

Worldwide evolution of mosquito resistance to chemical insecticides represents a major challenge for public health, and the future of vector control largely relies on the development of biological insecticides that can be used in combination with chemicals (integrated management), with the expectation that populations already resistant to chemicals will not become readily resistant to biological insecticides. However, little is known about the metabolic pathways affected by selection with chemical or biological insecticides. Here we show that Aedes aegypti, a laboratory mosquito strain selected with a biological insecticide (Bacillus thuringiensis israelensis, Bti) evolved increased transcription of many genes coding for endopeptidases while most genes coding for detoxification enzymes were under-expressed. By contrast, in strains selected with chemicals, genes encoding detoxification enzymes were mostly over-expressed. In all the resistant strains, genes involved in immune response were under-transcribed, suggesting that basal immunity might be a general adjustment variable to compensate metabolic costs caused by insecticide selection. Bioassays generally showed no evidence for an increased susceptibility of selected strains towards the other insecticide type, and all chemical-resistant strains were as susceptible to Bti as the unselected parent strain, which is a good premise for sustainable integrated management of mosquito populations resistant to chemicals.

Dengue, chikungunya and zika viruses are pathogens with an increasing global impact. In the absence of an approved vaccine or therapy, their management relies on controlling the mosquito vectors. But traditional controls are inadequate, and the range of invasive species such as Aedes albopictus (Asian Tiger Mosquito) is expanding. Genetically modified mosquitoes are being tested, but their use has encountered regulatory barriers and public opposition in some countries. Wolbachia bacteria can cause a form of conditional sterility, which can provide an alternative to genetic modification or irradiation. It is unknown however, whether openly released, artificially infected male Ae. albopictus can competitively mate and sterilize females at a level adequate to suppress a field population. Also, the unintended establishment of Wolbachia at the introduction site could result from horizontal transmission or inadvertent female release. In 2014, an Experimental Use Permit from the United States Environmental Protection Agency approved a pilot field trial in Lexington, Kentucky, USA. Here, we present data showing localized reduction of both egg hatch and adult female numbers. The artificial Wolbachia type was not observed to establish in the field. The results are discussed in relation to the applied use of Wolbachia-infected males as a biopesticide to suppress field populations of Ae. albopictus. PMID:27659038

Mosquito (Aedes togoi) larva has to float its siphon on the water surface to breathe air. To elucidate the floating mechanism of the siphon, morphological structures, especially the flap-like lobes and spiracle of the siphon, were observed. Wettability and dynamic behavior of the lobes on the water surface were also experimentally examined. The lobes formed a hollow cone shape under water and expanded on the water surface. The spiracle was located at the base of the cone. The lobes exhibited hydrophobic wettability. During floating process, the lobes were spread into a triangular shape in apical view, and the spiracle was exposed to air. When they were submerged in water, the lobes were folded into a cone shape to seal the spiracle and protect it from water penetration. These dynamic processes occurred at the water surface. In this study, the floating mechanism of the lobes for respiration was described based on the cross-sectional morphology of the lobes with the pressure induced by surface tension and the hydrostatic pressure proportional to depth. This study would be helpful in elucidating the behavior of mosquito larvae near the water surface and may facilitate the formulation of methods to control mosquito larvae. PMID:28216635

Full Text Available Abstract Background Aedes aegypti is arguably the most studied of all mosquito species in the laboratory and is the primary vector of both Dengue and Yellow Fever flaviviruses in the field. A large number of transcriptional studies have been made in the species and these usually report transcript quantities observed at a certain age or stage of development. However, circadian oscillation is an important characteristic of gene expression in many animals and plants, modulating both their physiology and behavior. Circadian gene expression in mosquito species has been previously reported but for only a few genes directly involved in the function of the molecular clock. Results Herein we analyze the transcription profiles of 21,494 messenger RNAs using an Ae. aegypti Agilent® microarray. Transcripts were quantified in adult female heads at 24 hours and then again at 72 hours and eight subsequent time points spaced four hours apart. We document circadian rhythms in multiple molecular pathways essential for growth, development, immune response, detoxification/pesticide resistance. Circadian rhythms were also noted in ribosomal protein genes used for normalization in reverse transcribed PCR (RT-PCR to determine transcript abundance. We report pervasive oscillations and intricate synchronization patterns relevant to all known biological pathways. Conclusion These results argue strongly that transcriptional analyses either need to be made over time periods rather than confining analyses to a single time point or development stage or exceptional care needs to be made to synchronize all mosquitoes to be analyzed and compared among treatment groups.

We report efficient germ-line transformation in the yellow fever mosquitoAedes aegypti accomplished using the piggyBac transposable element vector pBac[3xP3-EGFP afm]. Two transgenic lines were established and characterized; each contained the Vg-Defensin A transgene with strong eye-specific expression of the enhanced green fluorescent protein (EGFP) marker gene regulated by the artificial 3xP3 promoter. Southern blot hybridization and inverse PCR analyses of genomic DNA demonstrated a precise piggyBac-mediated, single copy insertion of the pBac[3xP3-EGFP afm,Vg-DefA] transposon in each transgenic line. For each line, genetic analysis confirmed stability and integrity of the entire transposon construct in the mosquito genome through the G2-G6 generations. Successful establishment of homozygous transgenic lines indicated that in both cases a non-lethal integration of the transposon into the mosquito genome had occurred. The 3xP3-EGFP marker was tested in mosquitoes with different genetic backgrounds. In white-eyed transgenic mosquitoes, the strong eye-specific expression of GFP was observed throughout all stages of development, starting from newly hatched first instar larvae to adults. A similar level and pattern of fluorescence was observed in red-eyed mosquitoes that were generated by crossing the 3xP3-EGFP transformants with the kh(w) white-eye mosquitoes transformed with the Drosophila cinnabar gene. Importantly, the utility of the 3xP3-EGFP, as marker gene for transformation of wild type mosquitoes, was demonstrated by strong eye-specific GFP expression in larval and pupal stages of black-eyed hybrids of the 3xP3-EGFP white-eye transformants and the wild type Rockefeller/UGAL strain. Finally, analysis of the Vg-DefA transgene expression in transformants from two established lines demonstrated strong blood-meal activation and fat-body-specific expression regulated by the Vg 1.8-kb 5' upstream region.

Lateralisation (i.e. functional and/or structural specialisations of left and right sides of the brain) of aggressive traits has been studied in a number of vertebrates, while evidence for invertebrates is scarce. Mosquito females display aggressive responses against undesired males, performing rejection kicks with the hind legs. In this research, we examined lateralisation of kicking behaviour in females of the Asian tiger mosquito, Aedes albopictus. We found a right-biased population-level lateralisation of kicking behaviour. Four repeated testing phases on mosquito females confirmed the preferential use of right legs. However, when left legs were used, the mean number of kicks per rejection event was not different to that performed with right legs. Both left and right kicking behaviour lead to successful displacement of undesired partners. This is the first report about behavioural lateralisation in mosquitoes.

Background The microbiota has been shown to play an important role in the biology of insects. In recent decades, significant efforts have been made to better understand the diversity of symbiotic bacteria associated with mosquitoes and assess their influence on pathogen transmission. Here, we report the bacterial composition found in field-caught Aedes albopictus populations by using culture-dependent methods. Results A total of 104 mosquito imagos (56 males and 48 females) were caught from four contrasting biotopes of Madagascar and their bacterial contents were screened by plating whole body homogenates on three different culture media. From 281 bacterial colony types obtained, amplified ribosomal DNA restriction analysis (ARDRA) showed they had 40 distinct ribotypes. Sequencing and BLAST analysis of the 16S rDNA genes responsible for each representative profile made it possible to identify 27 genera distributed in three major phyla. In female mosquitoes, bacterial isolates were mostly Proteobacteria (51.3%) followed by Firmicutes (30.3%) and Actinobacteria (18.4%). Conversely, Actinobacteria was the most abundant phylum in male mosquitoes (48%) followed by Proteobacteria (30.6%) and Firmicutes (20.4%). The relative abundance and composition of isolates also varied between sampling sites, ranging from 3 distinct families in Ankazobe to 8 in Tsimbazaza Park, and Toamasina and Ambohidratrimo. Pantoea was the most common genus in both females and males from all sampling sites, except for Ambohidratrimo. No differences in genome size were found between Pantoea isolates from mosquitoes and reference strains in pulse field gel electrophoresis. However, according to the numbers and sizes of plasmids, mosquito isolates clustered into three different groups with other strains isolated from insects but distinct from isolates from the environment. Conclusions The recent upsurge in research into the functional role of the insect microbiota prompts the interest to better

We identified species of copepods in natural mosquito breeding sites in the region of Soconusco, Chiapas, Mexico and compared their predatory ability on first instar larvae of Aedes aegypti and A. albopictus...

Mosquito coils are insecticides commonly used for protection against mosquitoes due to their toxic effects on mosquito populations. These effects on mosquitoes could induce the expression of metabolic enzymes in exposed populations as a counteractive measure. Cytochrome P450 family 4 (CYP4) are metabolic enzymes associated with a wide range of biological activities including insecticide resistance. In this study, the efficacies of three commercial mosquito coils with different pyrethroid active ingredients were assessed and their potential to induce the expression of CYP4 genes in Aedes albopictus analyzed by real-time quantitative PCR. Coils containing 0.3 % D-allethrin and 0.005 % metofluthrin exacted profound toxic effects on Ae. albopictus, inducing high mortalities (≥90 %) compared to the 0.2 % D-allethrin reference coil. CYP4H42 and CYP4H43 expressions were significantly higher in 0.3 % D-allethrin treated mosquitoes compared to the other treated populations. Short-term (KT50) exposure to mosquito coils induced significantly higher expression of both genes in 0.005 % metofluthrin exposed mosquitoes. These results suggest the evaluated products provided better protection than the reference coil; however, they also induced the expression of metabolic genes which could impact negatively on personal protection against mosquito.

Full Text Available Aedes aegypti is the major vector of yellow and dengue fevers. After 10 generations of adult selection, an A. aegypti strain (SP developed 1650-fold resistance to permethrin, which is one of the most widely used pyrethroid insecticides for mosquito control. SP larvae also developed 8790-fold resistance following selection of the adults. Prior to the selections, the frequencies of V1016G and F1534C mutations in domains II and III, respectively, of voltage-sensitive sodium channel (Vssc, the target site of pyrethroid insecticide were 0.44 and 0.56, respectively. In contrast, only G1016 alleles were present after two permethrin selections, indicating that G1016 can more contribute to the insensitivity of Vssc than C1534. In vivo metabolism studies showed that the SP strain excreted permethrin metabolites more rapidly than a susceptible SMK strain. Pretreatment with piperonyl butoxide caused strong inhibition of excretion of permethrin metabolites, suggesting that cytochrome P450 monooxygenases (P450s play an important role in resistance development. In vitro metabolism studies also indicated an association of P450s with resistance. Microarray analysis showed that multiple P450 genes were over expressed during the larval and adult stages in the SP strain. Following quantitative real time PCR, we focused on two P450 isoforms, CYP9M6 and CYP6BB2. Transcription levels of these P450s were well correlated with the rate of permethrin excretion and they were certainly capable of detoxifying permethrin to 4'-HO-permethrin. Over expression of CYP9M6 was partially due to gene amplification. There was no significant difference in the rate of permethrin reduction from cuticle between SP and SMK strains.

Full Text Available Objective: To evaluate the current knowledge level and information on Aedesmosquitoes and dengue among Penang resident in urban (Sungai Dua and sub- urban areas (Batu Maung. Methods: The questionnaire comprises of twenty-three questions, surveyed on 202 respondents regarding socio-demographic, the observation of general Aedes’ mosquito behaviour, dengue infection threat, preventive measures against mosquitoes and lifestyle (on usage of artificial light in changing the behaviour of Aedesmosquitoes. Results: The respondents in sub-urban Batu Maung showed less knowledge level regarding Aedesmosquitoes as compared to respondents in urban Sungai Dua. There was a positive association between biting frequency of mosquitoes with the lifestyle of respondents (in relation to the light use, the use of personal protection and fogging operations conducted by the Ministry of Health. There is a positive relationship between the knowledge of preferred mosquitoes resting place and the respondent aged between 17 to 24 (78.2% in sub-urban Batu Maung. Urban Sungai Dua respondents showed a significant relation between the use of lights and the resting places for Aedesmosquitoes. The preventive measures chosen by both sub-distinct areas against mosquitobites were significantly affected by the biting time and the mosquitoes biting frequency (P < 0.05. Conclusions: In this study, we concluded that there is a direct link between the knowledge of the Aedes biting frequency with dengue infection, human lifestyle on the light use and preventive measures against mosquitoes. However, the level of knowledge is poor on the subject of Aedesmosquitoes and dengue for both urban and sub-urban respondents.

Aedesaegypti has 2 genes encoding xanthine dehydrogenase (XDH). We analyzed XDH1 and XDH2 gene expression by real-time quantitative PCR in tissues from sugar- and blood-fed females. Differential XDH1 and XDH2 gene expression was observed in tissues dissected throughout a time course. We next exposed females to blood meals supplemented with allopurinol, a well-characterized XDH inhibitor. We also tested the effects of injecting double-stranded RNA (dsRNA) against XDH1, XDH2, or both. Disruption of XDH by allopurinol or XDH1 by RNA interference significantly affected mosquito survival, causing a disruption in blood digestion, excretion, oviposition, and reproduction. XDH1-deficient mosquitoes showed a persistence of serine proteases in the midgut at 48 h after blood feeding and a reduction in the uptake of vitellogenin by the ovaries. Surprisingly, analysis of the fat body from dsRNA-XDH1-injected mosquitoes fell into 2 groups: one group was characterized by a reduction of the XDH1 transcript, whereas the other group was characterized by an up-regulation of several transcripts including XDH1, glutamine synthetase, alanine aminotransferase, catalase, superoxide dismutase, ornithine decarboxylase, glutamate receptor, and ammonia transporter. Our data demonstrate that XDH1 plays an essential role and that XDH1 has the potential to be used as a metabolic target for Ae.aegypti vector control.-Isoe, J., Petchampai, N., Isoe, Y. E., Co, K., Mazzalupo, S., Scaraffia, P. Y. Xanthine dehydrogenase-1 silencing in Aedes aegypti mosquitoes promotes a blood feeding-induced adulticidal activity.

Full Text Available An investigation was carried out during April to December 2000 to study mosquito fauna and ecology in Guilan Prov¬ince of northern Iran. The mosquito larvae were collected by dipping method and larval habitat characteristics recorded ac¬cording to hydro-ecological features. In total, 3937 larvae of the genus Culex from 92 larval breeding sites were collected. Six spe¬cies of the genus Culex; Cx. mimeticus, Cx. pipiens, Cx. theileri, Cx. tritaeniorhynchus, Cx. hortensis, and Cx. terri¬tans were identified in the province and respectively comprised 10.3%, 47.2%, 2.2%, 31%, 6.5%, and 2.8% of the samples. Most of the larvae were collected from the natural habitats (75.6% such as river edges (6.5%, riverbed pools (28.2%, rain pools (47.8%, stream edges (9.4%, grasslands (1.9%, marshes (2.8%, and hoof-prints (3.4% and others from artificial habitats (24.4% including rice fields (32.1%, irrigation channels (7.1%, wells (16.4%, discarded concrete tubes (33.1%, dis¬carded tires (11.0%, and agricultural water-storage pools (0.3%. The ecology of Cx. pipiens and Cx. tritaeniorhynchus, which are the most prevalent species and potentially involved in the transmission of many pathogens to humans and domes¬ticated animals, must be extensively studied.

Full Text Available Introduction Aedes aegypti is responsible for the transmission of the dengue and yellow fever viruses. This study evaluated the effects of extracts from Cnidosculos phyllacanthus, Ricinus communis, and Coutarea hexandra on the developmental periods of A.aegypti larvae and pupae. Crude extracts of C. phyllacanthus and C. hexandra and oil from R. communis and C. phyllacanthus were used. Methods Bioassays of the larvicidal and pupicidal effects of these products at different concentrations and times of exposure were evaluated. The lethal and sublethal effects were determined using different concentrations in larvicidal tests. Mortality data were evaluated by Probit analysis to determine the LC50 and LC90 values. Results The vegetable oils from C. phyllacanthus and R. communis demonstrated greater efficiency for larval control with an LC50=0.28µl/mL and an LC90=1.48µl/mL and LC50=0.029µl/mL and a LC90=0.26µl/mL, respectively. In pupal tests toxic effects for all insects were verified after exposure to the products at significant LC50 and LC90 values for 24 and 48h. The effects of sublethal concentrations of C. phyllacanthus (oil were more effective on the insects. Conclusions The vegetables oils from C. phyllacanthus and R. communis demonstrated greater potential from the control of different developmental periods in the life cycle of this insect.

Since ancient times, plant products were used in various aspects. However, their use against pests decreased when chemical products became developed. Recently, concerns increased with respect to public health and environmental security requiring detection of natural products that may be used against insect pests. In this study, 41 plant extracts and 11 oil mixtures were evaluated against the yellow fever mosquito, Aedes aegypti (Linnaeus), the malaria vector, Anopheles stephensi (Liston), and the filariasis and encephalitis vector, Culex quinquefasciatus (Say) (Diptera: Culicidae) using the skin of human volunteers to find out the protection time and repellency. The five most effective oils were those of Litsea (Litsea cubeba), Cajeput (Melaleuca leucadendron), Niaouli (Melaleuca quinquenervia), Violet (Viola odorata), and Catnip (Nepeta cataria), which induced a protection time of 8 h at the maximum and a 100% repellency against all three species. This effect needs, however, a peculiar formulation to fix them on the human skin.

There is growing interest in applying the sterile insect technique (SIT) against mosquitoes. Mass production of mosquitoes for large-scale releases demands a huge amount of water. Yet, many arid and/or seasonally arid countries face the difficulties of acute water shortage, deterioration of water quality and environmental constraints. The re-use of water to rear successive generations of larvae is attractive as a way to reduce water usage and running costs, and help to make this control method viable. To determine whether dirty larval water was a suitable rearing medium for Anopheles arabiensis, in place of the 'clean' dechlorinated water routinely used, a series of three experiments was carried out to evaluate the effect of dirty water or mixed clean and dirty water on several parameters of insect quality. Batches of 100 fresh eggs were distributed in dirty water or added to clean water to test the effect of dirty water on egg hatching, whereas first-instar larvae were used to determine the effect on immature development time, pupation, adult emergence, body size, and longevity. Moreover, to assess the effect of dirty water on larval mortality, pupation rate, adult emergence, and longevity, L4 larvae collected after the tilting or larvae/pupae separation events were returned either to the dirty water or added to clean water. Results indicated that reusing dirty water or using a 50:50 mix of clean and dirty water did not affect egg hatching. Moreover, no difference was found in time to pupation, larval mortality or sex ratio when first-instar larvae were added to clean water, dirty water, or a 75:25, 50:50 or 25:75 mix of clean and dirty water and reared until emergence. When late-instar larvae were put back into their own rearing water, there was no effect on pupation rate, emergence rate or female longevity, though male longevity was reduced. When reared from first-instar larvae, however, dirty water decreased pupation rate, emergence rate, body size, and adult

Full Text Available The identification of arthropod bloodmeals is important in many epidemiological studies, as, the understanding of the life cycle of vectors and the patogens they transmit, as well as helping to define arthropods' control strategies. The precipitin test has been used for decades, but ELISA is slowly becoming more popular. To compare the two tests for sensitivity, specificity and accuracy to detect small insect bloodmeals, Aedes aegypti or Ae. fluviatilis mosquitoes were fed either on feline, canine or human hosts. Mosquitoes were frozen at 6, 12, 24, 48 or 72 h after feeding. Precipitin test showed better specificity and accuracy and ELISA test showed higher sensitivity. Better results with both tests were achieved when mosquitoes were frozen within 48 h from feeding.

Full Text Available BACKGROUND: Aedes aegypti and Aedes albopictus are potential vectors of chikungunya virus (CHIKV. The recent CHIKV outbreaks were caused by a new variant characterized by a mutation in the E1 glycoprotein gene (E1-226V which has favored a better transmissibility by Ae. albopictus. As Ae. albopictus tends to replace Ae. aegypti in many regions, one question remained: is Ae. albopictus as efficient as Ae. aegypti to transmit the variant E1-226V of CHIKV? METHODOLOGY AND FINDINGS: We infected orally both species with the variant E1-226V and estimated the infection, the viral dissemination, and the transmission rate by real time RT-PCR. Additionally, we used an in vitro assay to determine the amount of virus delivered by mosquitoes in their saliva. We found that Ae. aegypti as well as Ae. albopictus ensured a high replication of the virus which underwent an efficient dissemination as detectable in the salivary glands at day 2 post-infection (pi. Infectious CHIKV particles were delivered by salivary glands from day 2 with a maximum at day 6 pi for Ae. albopictus (10(3.3 PFU and day 7 pi for Ae. aegypti (10(2.5 PFU. CONCLUSIONS: Ae. albopictus is slightly more efficient than Ae. aegypti to transmit the variant E1-226V of CHIKV. These results will help to design an efficient vector control to limit transmission as soon as the first human cases are diagnosed.

In the mosquito midgut, luminal pH regulation and cellular ion transport processes are important for the digestion of food and maintenance of cellular homeostasis. pH regulation in the mosquito gut is affected by the vectorial movement of the principal ions including bicarbonate/carbonate and protons. As in all metazoans, mosquitoes employ the product of aerobic metabolism carbon dioxide in its bicarbonate/carbonate form as one of the major buffers of cellular and extracellular pH. The conversion of metabolic carbon dioxide to bicarbonate/carbonate is accomplished by a family of enzymes encoded by the carbonic anhydrase gene family. This study characterizes Aedes aegypti carbonic anhydrases using bioinformatic, molecular, and immunohistochemical methods. Our analyses show that there are fourteen Aedes aegypti carbonic anhydrase genes, two of which are expressed as splice variants. The carbonic anhydrases were classified as either integral membrane, peripheral membrane, mitochondrial, secreted, or soluble cytoplasmic proteins. Using polymerase chain reaction and Western blotting, one of the carbonic anhydrases, Aedes aegypti carbonic anhydrase 9, was analyzed and found in each life stage, male/female pupae, male/female adults, and in the female posterior midgut. Next, carbonic anhydrase 9 was analyzed in larvae and adults using confocal microscopy and was detected in the midgut regions. According to our analyses, carbonic anhydrase 9 is a soluble cytoplasmic enzyme found in the alimentary canal of larvae and adults and is expressed throughout the life cycle of the mosquito. Based on previous physiological analyses of adults and larvae, it appears AeCA9 is one of the major carbonic anhydrases involved in producing bicarbonate/carbonate which is involved in pH regulation and ion transport processes in the alimentary canal. Detailed understanding of the molecular bases of ion homeostasis in mosquitoes will provide targets for novel mosquito control strategies into the

In the mosquito midgut, luminal pH regulation and cellular ion transport processes are important for the digestion of food and maintenance of cellular homeostasis. pH regulation in the mosquito gut is affected by the vectorial movement of the principal ions including bicarbonate/carbonate and protons. As in all metazoans, mosquitoes employ the product of aerobic metabolism carbon dioxide in its bicarbonate/carbonate form as one of the major buffers of cellular and extracellular pH. The conversion of metabolic carbon dioxide to bicarbonate/carbonate is accomplished by a family of enzymes encoded by the carbonic anhydrase gene family. This study characterizes Aedes aegypti carbonic anhydrases using bioinformatic, molecular, and immunohistochemical methods. Our analyses show that there are fourteen Aedes aegypti carbonic anhydrase genes, two of which are expressed as splice variants. The carbonic anhydrases were classified as either integral membrane, peripheral membrane, mitochondrial, secreted, or soluble cytoplasmic proteins. Using polymerase chain reaction and Western blotting, one of the carbonic anhydrases, Aedes aegypti carbonic anhydrase 9, was analyzed and found in each life stage, male/female pupae, male/female adults, and in the female posterior midgut. Next, carbonic anhydrase 9 was analyzed in larvae and adults using confocal microscopy and was detected in the midgut regions. According to our analyses, carbonic anhydrase 9 is a soluble cytoplasmic enzyme found in the alimentary canal of larvae and adults and is expressed throughout the life cycle of the mosquito. Based on previous physiological analyses of adults and larvae, it appears AeCA9 is one of the major carbonic anhydrases involved in producing bicarbonate/carbonate which is involved in pH regulation and ion transport processes in the alimentary canal. Detailed understanding of the molecular bases of ion homeostasis in mosquitoes will provide targets for novel mosquito control strategies into the

Aedes aegypti is an anautogenous mosquito that must blood feed on a vertebrate host to produce and lay a clutch of eggs. The rockpool mosquito, Georgecraigius atropalpus, is related to A. aegypti but is a facultatively autogenous species that produces its first clutch of eggs shortly after emerging without blood feeding. Consumption of a blood meal by A. aegypti triggers the release of ovary ecdysteroidogenic hormone (OEH) and insulin-like peptide 3 (ILP3) from the brain, which stimulate egg formation. OEH and ILP3 also stimulate egg formation in G. atropalpus but are released at eclosion independently of blood feeding. These results collectively suggest that blood meal dependent release of OEH and ILP3 is one factor that prevents A. aegypti from reproducing autogenously. Here, we examined two other factors that potentially inhibit autogeny in A. aegypti: teneral nutrient reserves and the ability of OEH and ILP3 to stimulate egg formation in the absence of blood feeding. Measures of nutrient reserves showed that newly emerged A. aegypti females had similar wet weights but significantly lower protein and glycogen reserves than G. atropalpus females when larvae were reared under identical conditions. OEH stimulated non-blood fed A. aegypti females to produce ecdysteroid hormone and package yolk into oocytes more strongly than ILP3. OEH also reduced host seeking and blood feeding behavior, yet females produced few mature eggs. Overall, our results indicate that multiple factors prevent A. aegypti from reproducing autogenously.

Bursicon is a neuropeptide hormone consisting of two cystine-knot proteins (burs α and burs β), responsible for cuticle tanning and other developmental processes in insects. Recent studies show that each bursicon subunit forms homodimers that induce prophylactic immunity in Drosophila melanogaster. Here, we investigated the hypothesis that bursicon homodimers act in prophylactic immunity in insects, and possibly arthropods, generally, using the mosquito, Aedes aegypti. We found that burs α and burs β are expressed in larvae, pupae and newly emerged adults. Treating newly emerged Ae. aegypti and D. melanogaster adults with recombinant bursicon (r-bursicon) heterodimer led to cuticle tanning in both species. Treating larvae and adults with r-bursicon homodimers led to up-regulation of five anti-microbial peptide (AMP) genes, noting the possibility that bursicon heterodimers also lead to up-regulation of these genes can not been excluded. The induced AMPs effectively suppressed the growth of bacteria in vitro. RNAi knock-down of the transcriptional factor Relish2 abolished the influence of r-bursicon homodimers on AMP production. We infer the bursicon homodimers induce expression of AMP genes via Relish2 in Ae. aegypti, as prophylactic immunity to protect mosquitoes during the vulnerable stages of each molt. PMID:28225068

The value of using plant volatiles as attractants for trapping and spatial repellents to protect hosts against mosquitoes has been widely recognized. The current study characterized behavioral responses of Aedes albopictus (Skuse) (Diptera: Culicidae) to different concentrations, ranging from 6 to 96%, of several common floral odorous compounds, including linalool, geraniol, citronellal, eugenol, anisaldehyde, and citral, using a wind tunnel olfactometer system. The results indicated that female mosquitoes reacted differently to different concentrations of the tested compounds, and the reactions also were different when those chemicals were tested alone or in the presence of human host odor. When tested alone, anisaldehyde was attractive at all tested concentrations, eugenol was attractive only at concentrations of 48-96%, while citronellal, linalool, citral, and geraniol were attractive at lower concentrations and repellent at higher concentrations. When tested in the presence of a human host, all compounds except for anisaldehyde at all tested concentrations showed host-seeking inhibition to certain degrees. Based on the results, it was concluded that anisaldehyde was effective in attracting Ae. albopictus when used alone but could also remarkably inhibit the host-seeking ability at a concentration of 96%, while citral, geraniol, linalool, citronellal, and eugenol are suitable as spatial repellents.

Bursicon is a neuropeptide hormone consisting of two cystine-knot proteins (burs α and burs β), responsible for cuticle tanning and other developmental processes in insects. Recent studies show that each bursicon subunit forms homodimers that induce prophylactic immunity in Drosophila melanogaster. Here, we investigated the hypothesis that bursicon homodimers act in prophylactic immunity in insects, and possibly arthropods, generally, using the mosquito, Aedes aegypti. We found that burs α and burs β are expressed in larvae, pupae and newly emerged adults. Treating newly emerged Ae. aegypti and D. melanogaster adults with recombinant bursicon (r-bursicon) heterodimer led to cuticle tanning in both species. Treating larvae and adults with r-bursicon homodimers led to up-regulation of five anti-microbial peptide (AMP) genes, noting the possibility that bursicon heterodimers also lead to up-regulation of these genes can not been excluded. The induced AMPs effectively suppressed the growth of bacteria in vitro. RNAi knock-down of the transcriptional factor Relish2 abolished the influence of r-bursicon homodimers on AMP production. We infer the bursicon homodimers induce expression of AMP genes via Relish2 in Ae. aegypti, as prophylactic immunity to protect mosquitoes during the vulnerable stages of each molt.

Background In the model system Drosophila melanogaster, doublesex (dsx) is the double-switch gene at the bottom of the somatic sex determination cascade that determines the differentiation of sexually dimorphic traits. Homologues of dsx are functionally conserved in various dipteran species, including the malaria vector Anopheles gambiae. They show a striking conservation of sex-specific regulation, based on alternative splicing, and of the encoded sex-specific proteins, which are transcriptional regulators of downstream terminal genes that influence sexual differentiation of cells, tissues and organs. Results In this work, we report on the molecular characterization of the dsx homologue in the dengue and yellow fever vector Aedes aegypti (Aeadsx). Aeadsx produces sex-specific transcripts by alternative splicing, which encode isoforms with a high degree of identity to Anopheles gambiae and Drosophila melanogaster homologues. Interestingly, Aeadsx produces an additional novel female-specific splicing variant. Genomic comparative analyses between the Aedes and Anopheles dsx genes revealed a partial conservation of the exon organization and extensive divergence in the intron lengths. An expression analysis showed that Aeadsx transcripts were present from early stages of development and that sex-specific regulation starts at least from late larval stages. The analysis of the female-specific untranslated region (UTR) led to the identification of putative regulatory cis-elements potentially involved in the sex-specific splicing regulation. The Aedes dsx sex-specific splicing regulation seems to be more complex with the respect of other dipteran species, suggesting slightly novel evolutionary trajectories for its regulation and hence for the recruitment of upstream splicing regulators. Conclusions This study led to uncover the molecular evolution of Aedes aegypti dsx splicing regulation with the respect of the more closely related Culicidae Anopheles gambiae orthologue

Full Text Available Abstract Background In the model system Drosophila melanogaster, doublesex (dsx is the double-switch gene at the bottom of the somatic sex determination cascade that determines the differentiation of sexually dimorphic traits. Homologues of dsx are functionally conserved in various dipteran species, including the malaria vector Anopheles gambiae. They show a striking conservation of sex-specific regulation, based on alternative splicing, and of the encoded sex-specific proteins, which are transcriptional regulators of downstream terminal genes that influence sexual differentiation of cells, tissues and organs. Results In this work, we report on the molecular characterization of the dsx homologue in the dengue and yellow fever vector Aedes aegypti (Aeadsx. Aeadsx produces sex-specific transcripts by alternative splicing, which encode isoforms with a high degree of identity to Anopheles gambiae and Drosophila melanogaster homologues. Interestingly, Aeadsx produces an additional novel female-specific splicing variant. Genomic comparative analyses between the Aedes and Anopheles dsx genes revealed a partial conservation of the exon organization and extensive divergence in the intron lengths. An expression analysis showed that Aeadsx transcripts were present from early stages of development and that sex-specific regulation starts at least from late larval stages. The analysis of the female-specific untranslated region (UTR led to the identification of putative regulatory cis-elements potentially involved in the sex-specific splicing regulation. The Aedes dsx sex-specific splicing regulation seems to be more complex with the respect of other dipteran species, suggesting slightly novel evolutionary trajectories for its regulation and hence for the recruitment of upstream splicing regulators. Conclusions This study led to uncover the molecular evolution of Aedes aegypti dsx splicing regulation with the respect of the more closely related Culicidae

We report here the first evidence of vertical transmission of Aedes flavivirus (AEFV) and its first isolation in the Western Hemisphere. AEFV strain SPFLD-MO-2011-MP6 was isolated in C6/36 cells from a pool of male Aedes albopictus mosquitoes that were reared to adults from larvae collected in southwest Missouri, USA, in 2011. Electron micrographs of the virus showed virions of approximately 45 nm in diameter with morphological characteristics associated with flaviviruses. The genomic sequence demonstrated that AEFV-SPFLD-MO-2011-MP6 shares a high degree of nucleotide and amino acid sequence identity with the AEFV Narita-21 strain, isolated in Japan in 2003. Intracerebral inoculation of newborn mice with the virus failed to produce observable illness or death and the virus did not replicate in vertebrate cells, consistent with a lack of vertebrate host range. - Highlights: ► The first report of Aedes flavivirus (AEFV) in the Western Hemisphere. ► The first evidence of vertical transmission of AEFV in mosquitoes. ► The first electron micrograph of AEFV. ► The first attempt to infect animals with AEFV.

Three gustatory receptor neurons were characterized for contact chemoreceptive sensilla on the labella of female yellow-fever mosquitoes, Aedes aegypti. The neuron with the smallest amplitude spike responded to the feeding deterrent, quinine, as well as N, N-diethyl-3-methylbenzamide and other insect repellents. Two other neurons with differing spikes responded to salt (NaCl) and sucrose. This is the first report of a gustatory receptor neuron specific for insect repellents in mosquitoes and may provide a tool for screening chemicals to discover novel or improved feeding deterrents and repellents for use in the management of arthropod disease vectors.

Full Text Available Abstract Background Aedes aegypti mosquitoes are the main vectors of dengue viruses to humans. Understanding their biology and interactions with the pathogen are prerequisites for development of dengue transmission control strategies. Mosquito salivary glands are organs involved directly in pathogen transmission to vertebrate hosts. Information on the spatial distribution of gene expression in these organs is expected to assist in the development of novel disease control strategies, including those that entail the release of transgenic mosquitoes with impaired vector competence. Results We report here the hybridization in situ patterns of 30 transcripts expressed in the salivary glands of adult Ae. aegypti females. Distinct spatial accumulation patterns were identified. The products of twelve genes are localized exclusively in the proximal-lateral lobes. Among these, three accumulate preferentially in the most anterior portion of the proximal-lateral lobe. This pattern revealed a salivary gland cell type previously undescribed in Ae. aegypti, which was validated by transmission electron microscopy. Five distinct gene products accumulate in the distal-lateral lobes and another five localize in the medial lobe. Seven transcripts are found in the distal-lateral and medial lobes. The transcriptional product of one gene accumulates in proximal- and distal-lateral lobes. Seven genes analyzed by quantitative PCR are expressed constitutively. The most abundant salivary gland transcripts are those localized within the proximal-lateral lobes, while previous work has shown that the distal-lateral lobes are the most active in protein synthesis. This incongruity suggests a role for translational regulation in mosquito saliva production. Conclusions Transgenic mosquitoes with reduced vector competence have been proposed as tools for the control of dengue virus transmission. Expression of anti-dengue effector molecules in the distal-lateral lobes of Ae

Full Text Available Abstract Background In the past two decades the east African highlands have experienced several major malaria epidemics. Currently there is a renewed interest in exploring the possibility of anopheline larval control through environmental management or larvicide as an additional means of reducing malaria transmission in Africa. This study examined the landscape determinants of anopheline mosquitolarval habitats and usefulness of remote sensing in identifying these habitats in western Kenya highlands. Methods Panchromatic aerial photos, Ikonos and Landsat Thematic Mapper 7 satellite images were acquired for a study area in Kakamega, western Kenya. Supervised classification of land-use and land-cover and visual identification of aquatic habitats were conducted. Ground survey of all aquatic habitats was conducted in the dry and rainy seasons in 2003. All habitats positive for anopheline larvae were identified. The retrieved data from the remote sensors were compared to the ground results on aquatic habitats and land-use. The probability of finding aquatic habitats and habitats with Anopheles larvae were modelled based on the digital elevation model and land-use types. Results The misclassification rate of land-cover types was 10.8% based on Ikonos imagery, 22.6% for panchromatic aerial photos and 39.2% for Landsat TM 7 imagery. The Ikonos image identified 40.6% of aquatic habitats, aerial photos identified 10.6%, and Landsate TM 7 image identified 0%. Computer models based on topographic features and land-cover information obtained from the Ikonos image yielded a misclassification rate of 20.3–22.7% for aquatic habitats, and 18.1–25.1% for anopheline-positive larval habitats. Conclusion One-metre spatial resolution Ikonos images combined with computer modelling based on topographic land-cover features are useful tools for identification of anopheline larval habitats, and they can be used to assist to malaria vector control in western Kenya

Full Text Available BACKGROUND: The fat body is the main organ of intermediary metabolism in insects and the principal source of hemolymph proteins. As part of our ongoing efforts to understand mosquito fat body physiology and to identify novel targets for insect control, we have conducted a transcriptome analysis of the fat body of Aedes aegypti before and in response to blood feeding. RESULTS: We created two fat body non-normalized EST libraries, one from mosquito fat bodies non-blood fed (NBF and another from mosquitoes 24 hrs post-blood meal (PBM. 454 pyrosequencing of the non-normalized libraries resulted in 204,578 useable reads from the NBF sample and 323,474 useable reads from the PBM sample. Alignment of reads to the existing reference Ae. aegypti transcript libraries for analysis of differential expression between NBF and PBM samples revealed 116,912 and 115,051 matches, respectively. De novo assembly of the reads from the NBF sample resulted in 15,456 contigs, and assembly of the reads from the PBM sample resulted in 15,010 contigs. Collectively, 123 novel transcripts were identified within these contigs. Prominently expressed transcripts in the NBF fat body library were represented by transcripts encoding ribosomal proteins. Thirty-five point four percent of all reads in the PBM library were represented by transcripts that encode yolk proteins. The most highly expressed were transcripts encoding members of the cathepsin b, vitellogenin, vitellogenic carboxypeptidase, and vitelline membrane protein families. CONCLUSION: The two fat body transcriptomes were considerably different from each other in terms of transcript expression in terms of abundances of transcripts and genes expressed. They reflect the physiological shift of the pre-feeding fat body from a resting state to vitellogenic gene expression after feeding.

The PIWI-interacting RNA (piRNA) pathway is essential for transposon silencing in many model organisms. Its remarkable efficiency relies on a sophisticated amplification mechanism known as the ping-pong loop. In Alphavirus-infected Aedesmosquitoes, piRNAs with sequence features that suggest ping-pong-dependent biogenesis are produced from viral RNA. The PIWI family in Aedesmosquitoes is expanded when compared to other model organisms, raising the possibility that individual PIWI proteins have functionally diversified in these insects. Here, we show that Piwi5 and Ago3, but none of the other PIWI family members, are essential for piRNA biogenesis from Sindbis virus RNA in infected Aedes aegypti cells. In contrast, the production of piRNAs from transposons relies on a more versatile set of PIWI proteins, some of which do not contribute to viral piRNA biogenesis. These results indicate that functional specialization allows distinct mosquito PIWI proteins to process RNA from different endogenous and exogenous sources.

The PIWI-interacting RNA (piRNA) pathway is essential for transposon silencing in many model organisms. Its remarkable efficiency relies on a sophisticated amplification mechanism known as the ping-pong loop. In Alphavirus-infected Aedesmosquitoes, piRNAs with sequence features that suggest ping-pong-dependent biogenesis are produced from viral RNA. The PIWI family in Aedesmosquitoes is expanded when compared to other model organisms, raising the possibility that individual PIWI proteins have functionally diversified in these insects. Here, we show that Piwi5 and Ago3, but none of the other PIWI family members, are essential for piRNA biogenesis from Sindbis virus RNA in infected Aedes aegypti cells. In contrast, the production of piRNAs from transposons relies on a more versatile set of PIWI proteins, some of which do not contribute to viral piRNA biogenesis. These results indicate that functional specialization allows distinct mosquito PIWI proteins to process RNA from different endogenous and exogenous sources. PMID:26068474

The Aedes aegypti vector for dengue virus (DENV) has been reported in urban and periurban areas. The information about DENV circulation in mosquitoes in Colombian rural areas is limited, so we aimed to evaluate the presence of DENV in Ae. aegypti females caught in rural locations of two Colombian municipalities, Anapoima and La Mesa. Mosquitoes from 497 rural households in 44 different rural settlements were collected. Pools of about 20 Ae. aegypti females were processed for DENV serotype detection. DENV in mosquitoes was detected in 74% of the analysed settlements with a pool positivity rate of 62%. The estimated individual mosquito infection rate was 4.12% and the minimum infection rate was 33.3/1,000 mosquitoes. All four serotypes were detected; the most frequent being DENV-2 (50%) and DENV-1 (35%). Two-three serotypes were detected simultaneously in separate pools. This is the first report on the co-occurrence of natural DENV infection of mosquitoes in Colombian rural areas. The findings are important for understanding dengue transmission and planning control strategies. A potential latent virus reservoir in rural areas could spill over to urban areas during population movements. Detecting DENV in wild-caught adult mosquitoes should be included in the development of dengue epidemic forecasting models.

We determined the feasibility of using the BG-Sentinel™ mosquito trap (BGS) as the pull component in a push-pull strategy to reduce indoor biting by Aedes aegypti. This included evaluating varying numbers of traps (1-4) and mosquito release numbers (10, 25, 50, 100, 150, 200, and 250) on recapture rates under screen house conditions. Based on these variations in trap and mosquito numbers, release intervals were rotated through a completely randomized design with environmental factors (temperature, relative humidity, and light intensity) and monitored throughout each experiment. Data from four sampling time points (05:30, 09:30, 13:30, and 17:30) indicate a recapture range among treatments of 66-98%. Furthermore, 2-3 traps were as effective in recapturing mosquitoes as 4 traps for all mosquito release numbers. Time trends indicate Day 1 (the day the mosquitoes were released) as the "impact period" for recapture with peak numbers of marked mosquitoes collected at 09:30 or 4 h post-release. Information from this study will be used to guide the configuration of the BGS trap component of a push-pull vector control strategy currently in the proof-of-concept stage of development in Thailand and Peru.

Full Text Available The Aedes aegypti vector for dengue virus (DENV has been reported in urban and periurban areas. The information about DENV circulation in mosquitoes in Colombian rural areas is limited, so we aimed to evaluate the presence of DENV in Ae. aegypti females caught in rural locations of two Colombian municipalities, Anapoima and La Mesa. Mosquitoes from 497 rural households in 44 different rural settlements were collected. Pools of about 20 Ae. aegypti females were processed for DENV serotype detection. DENV in mosquitoes was detected in 74% of the analysed settlements with a pool positivity rate of 62%. The estimated individual mosquito infection rate was 4.12% and the minimum infection rate was 33.3/1,000 mosquitoes. All four serotypes were detected; the most frequent being DENV-2 (50% and DENV-1 (35%. Two-three serotypes were detected simultaneously in separate pools. This is the first report on the co-occurrence of natural DENV infection of mosquitoes in Colombian rural areas. The findings are important for understanding dengue transmission and planning control strategies. A potential latent virus reservoir in rural areas could spill over to urban areas during population movements. Detecting DENV in wild-caught adult mosquitoes should be included in the development of dengue epidemic forecasting models.

A fundamental understanding of the spatial distribution and ecology of mosquito larvae is essential for effective vector control intervention strategies. In this study, data-driven decision tree models, generalized linear models and ordination analysis were used to identify the most important biotic and abiotic factors that affect the occurrence and abundance of mosquito larvae in Southwest Ethiopia. In total, 220 samples were taken at 180 sampling locations during the years 2010 and 2012. Sampling sites were characterized based on physical, chemical and biological attributes. The predictive performance of decision tree models was evaluated based on correctly classified instances (CCI), Cohen's kappa statistic (κ) and the determination coefficient (R2). A conditional analysis was performed on the regression tree models to test the relation between key environmental and biological parameters and the abundance of mosquito larvae. The decision tree model developed for anopheline larvae showed a good model performance (CCI = 84 ± 2%, and κ = 0.66 ± 0.04), indicating that the genus has clear habitat requirements. Anopheline mosquito larvae showed a widespread distribution and especially occurred in small human-made aquatic habitats. Water temperature, canopy cover, emergent vegetation cover, and presence of predators and competitors were found to be the main variables determining the abundance and distribution of anopheline larvae. In contrast, anopheline mosquito larvae were found to be less prominently present in permanent larval habitats. This could be attributed to the high abundance and diversity of natural predators and competitors suppressing the mosquito population densities. The findings of this study suggest that targeting smaller human-made aquatic habitats could result in effective larval control of anopheline mosquitoes in the study area. Controlling the occurrence of mosquito larvae via drainage of permanent wetlands may not be a good management

The spatial distribution of Aedes aegypti and Aedes albopictus in environmental and geographical zones, e.g., urban-rural, coastal-mountainous, and north-south, was investigated throughout Vietnam. Immature stages were collected from used tires along roads. The effects of regions, seasons, and the degree of urbanization on the density and the frequency were statistically analyzed. Aedes aegypti predominated in the southern and central regions, while Ae. albopictus predominated in the northern region, which may be related to climatic conditions (temperature and rainfall). Larval collection from used tires may be suitable to assess rapidly the current distribution of dengue mosquitoes for estimating health risks and implementing vector control measures.

The dengue and yellow fever mosquito, Aedes aegypti, contributes significantly to global disease burden. Genetic study of Aedes aegypti is essential to understanding its evolutionary history, competence as a disease vector, and the effects and efficacy of vector control methods. The prevalence of repeats and transposable elements in the Aedes aegypti genome complicates marker development and makes genome-wide genetic study challenging. To overcome these challenges, we developed a high-throughput genotyping chip, Axiom_aegypti1. This chip screens for 50,000 single-nucleotide polymorphisms present in Aedes aegypti populations from around the world. The array currently used genotypes 96 samples simultaneously. To ensure that these markers satisfy assumptions commonly made in many genetic analyses, we tested for Mendelian inheritance and linkage disequilibrium in laboratory crosses and a wild population, respectively. We have validated more than 25,000 of these markers to date, and expect this number to increase with more sampling. We also present evidence of the chip's efficacy in distinguishing populations throughout the world. The markers on this chip are ideal for applications ranging from population genetics to genome-wide association studies. This tool makes rapid, cost-effective, and comparable genotype data attainable to diverse sets of Aedes aegypti researchers, from those interested in potential range shifts due to climate change to those characterizing the genetic underpinnings of its competence to transmit disease.

Competition experiments estimating the relative effects of inter- and intraspecific competition can help to resolve whether interspecific competition results in coexistence or exclusion. For mosquitoes, most such experiments have focused on invasive Aedes albopictus and its interactions with resident Aedes. A meta-analysis of such experiments tested whether the effect of interspecific competition is greater than, less than, or equal to that of intraspecific competition, and whether competitive outcomes are dependent on food quality. For A. albopictus and A. aegypti, there was significant context dependence, with interspecific competitive advantage for A. albopictus with low food quality, and competitive equivalence with high food quality. Meta-analysis of survivorship yielded more significant effects than did estimated rate of increase. Competitive effects and competitive responses of each species yielded similar results. This meta-analysis suggests competitive exclusion of A. aegypti by A. albopictus, and is thus consistent with field sampling, qualitative reviews, and interpretations from individual publications. For A. albopictus and A. triseriatus, most results indicated competitive equivalence and no context dependence, and are thus contrary to previous qualitative reviews and to interpretations from individual publications. For both pairs of species, published results suitable for meta-analysis remain scarce, and better experimental designs and improved analysis and reporting of statistical results are needed. Greater emphasis needs to be placed on estimating species' inter- and intra-specific competitive effects, rather than the more common, but theoretically less interesting, competitive responses. Experiments without low-density controls (i.e., replacement series) are inadequate for comparing competitive effects and responses.

Full Text Available Selection of oviposition sites by gravid females is a critical behavioral step in the reproductive cycle of Anopheles coluzzii, which is one of the principal Afrotropical malaria vector mosquitoes. Several studies suggest this decision is mediated by semiochemicals associated with potential oviposition sites. To better understand the chemosensory basis of this behavior and identify compounds that can modulate oviposition, we examined the generally held hypothesis that suboptimal larval habitats give rise to semiochemicals that negatively influence the oviposition preference of gravid females. Dual-choice bioassays indicated that oviposition sites conditioned in this manner do indeed foster significant and concentration dependent aversive effects on the oviposition site selection of gravid females. Headspace analyses derived from aversive habitats consistently noted the presence of dimethyl disulfide (DMDS, dimethyl trisulfide (DMTS and 6-methyl-5-hepten-2-one (sulcatone each of which unitarily affected An. coluzzii oviposition preference. Electrophysiological assays across the antennae, maxillary palp, and labellum of gravid An. coluzzii revealed differential responses to these semiochemicals. Taken together, these findings validate the hypothesis in question and suggest that suboptimal environments for An. coluzzii larval development results in the release of DMDS, DMTS and sulcatone that impact the response valence of gravid females.

To determine the distribution of Aedes aegypti in Ciudad Juárez, Mexico and evaluate it as a carrier of the dengue virus. Mosquitoes were collected using CDC minitraps. Nearest neighbor and K-function were used as geospatial tools. The chi-square test was utilized to evaluate the association between the presence of the vector and sociodemographic variables. Evidence of infection was detected by RT-PCR. A total of 122 female mosquitoes were captured. A tendency in the cluster distribution (R= -1.2995, p= 0.05) of the mosquito was shown up to 4000 m but none of the sociodemographic variables showed significant associations. Seven of the pools tested were positive for DEN-2, ten were positive for DEN- 3, and seven for both serotypes. This is the first report on the presence of Aedes aegypti mosquitoes infected with dengue in the region, which will enable the promotion of monitoring in order to reduce the probability of occurrence of the disease among the border population.

A study was conducted in the city of Salto, Uruguay, to identify mosquito-producing containers, the spatial distribution of mosquitoes and the relationship between the different population indices of Aedes aegypti. On each of 312 premises visited, water-filled containers and immature Ae. aegypti mosquitoes were identified. The containers were counted and classified into six categories. Pupae per person and Stegomyia indices were calculated. Pupae per person were represented spatially. The number of each type of container and number of mosquitoes in each were analyzed and compared, and their spatial distribution was analyzed. No significant differences in the number of the different types of containers with mosquitoes or in the number of mosquitoes in each were found. The distribution of the containers with mosquito was random and the distribution of mosquitoes by type of container was aggregated or highly aggregated. PMID:23128295

Dried male inflorescences of breadfruit ( Artocarpus altilis , Moraceae) are burned in communities throughout Oceania to repel flying insects, including mosquitoes. This study was conducted to identify chemicals responsible for mosquito deterrence. Various crude extracts were evaluated, and the most active, the hydrodistillate, was used for bioassay-guided fractionation. The hydrodistillate and all fractions displayed significant deterrent activity. Exploratory GC-MS analysis revealed more than 100 distinctive peaks, and more than 30 compounds were putatively identified, including a mixture of terpenes, aldehydes, fatty acids, and aromatics. A systematic bioassay-directed study using adult Aedes aegypti females identified capric, undecanoic, and lauric acid as primary deterrent constituents. A synthetic mixture of fatty acids present in the most active fraction and individual fatty acids were all significantly more active than N,N-diethyl-m-toluamide (DEET). These results provide support for this traditional practice and indicate the potential of male breadfruit flowers and fatty acids as mosquito repellents.

Objective:To determine the adulticidal activity of hexane, benzene, chloroform, ethyl acetate and methanol leaf and seed extracts of Delonix elata (D. elata) against Aedes aegypti (Ae. aegypti). Methods:The bioassay was conducted in an experimental kit consisting of two cylindrical plastic tubes both measuring 125 mm×44 mm following the WHO method;mortality of the mosquitoes was recorded after 24 h. Results:The adulticidal activity of plant leaf and seed extracts showed moderate toxic effect on the adult mosquitoes after 24 h of exposure period. However, the highest adulticidal activity was observed in the leaf methanol extract of D. elata against Ae. aegypti with the LC50 and LC90 values 162.87 and 309.32 mg/L, respectively. Conclusions:From this result, it can be concluded the crude extract of D. elata was an excellent potential for controlling Ae. aegypti mosquitoes.

Full Text Available Zika virus (ZIKV and chikungunya virus (CHIKV are highly pathogenic arthropod-borne viruses that are currently a serious health burden in the Americas, and elsewhere in the world. ZIKV and CHIKV co-circulate in the same geographical regions and are mainly transmitted by Aedes aegypti mosquitoes. There is a growing number of case reports of ZIKV and CHIKV co-infections in humans, but it is uncertain whether co-infection occurs via single or multiple mosquito bites. Here we investigate the potential of Ae. aegypti mosquitoes to transmit both ZIKV and CHIKV in one bite, and we assess the consequences of co-infection on vector competence.First, growth curves indicated that co-infection with CHIKV negatively affects ZIKV production in mammalian, but not in mosquito cells. Next, Ae. aegypti mosquitoes were infected with ZIKV, CHIKV, or co-infected via an infectious blood meal or intrathoracic injections. Infection and transmission rates, as well as viral titers of positive mosquitoes, were determined at 14 days after blood meal or 7 days after injection. Saliva and bodies of (co-infected mosquitoes were scored concurrently for the presence of ZIKV and/or CHIKV using a dual-colour immunofluorescence assay. The results show that orally exposed Ae. aegypti mosquitoes are highly competent, with transmission rates of up to 73% for ZIKV, 21% for CHIKV, and 12% of mosquitoes transmitting both viruses in one bite. However, simultaneous oral exposure to both viruses did not change infection and transmission rates compared to exposure to a single virus. Intrathoracic injections indicate that the selected strain of Ae. aegypti has a strong salivary gland barrier for CHIKV, but a less profound barrier for ZIKV.This study shows that Ae. aegypti can transmit both ZIKV and CHIKV via a single bite. Furthermore, co-infection of ZIKV and CHIKV does not influence the vector competence of Ae. aegypti.

Zika virus (ZIKV) and chikungunya virus (CHIKV) are highly pathogenic arthropod-borne viruses that are currently a serious health burden in the Americas, and elsewhere in the world. ZIKV and CHIKV co-circulate in the same geographical regions and are mainly transmitted by Aedes aegypti mosquitoes. There is a growing number of case reports of ZIKV and CHIKV co-infections in humans, but it is uncertain whether co-infection occurs via single or multiple mosquito bites. Here we investigate the potential of Ae. aegypti mosquitoes to transmit both ZIKV and CHIKV in one bite, and we assess the consequences of co-infection on vector competence. First, growth curves indicated that co-infection with CHIKV negatively affects ZIKV production in mammalian, but not in mosquito cells. Next, Ae. aegypti mosquitoes were infected with ZIKV, CHIKV, or co-infected via an infectious blood meal or intrathoracic injections. Infection and transmission rates, as well as viral titers of positive mosquitoes, were determined at 14 days after blood meal or 7 days after injection. Saliva and bodies of (co-)infected mosquitoes were scored concurrently for the presence of ZIKV and/or CHIKV using a dual-colour immunofluorescence assay. The results show that orally exposed Ae. aegypti mosquitoes are highly competent, with transmission rates of up to 73% for ZIKV, 21% for CHIKV, and 12% of mosquitoes transmitting both viruses in one bite. However, simultaneous oral exposure to both viruses did not change infection and transmission rates compared to exposure to a single virus. Intrathoracic injections indicate that the selected strain of Ae. aegypti has a strong salivary gland barrier for CHIKV, but a less profound barrier for ZIKV. This study shows that Ae. aegypti can transmit both ZIKV and CHIKV via a single bite. Furthermore, co-infection of ZIKV and CHIKV does not influence the vector competence of Ae. aegypti.

Full Text Available The mosquitoAedes aegypti (L. is a major vector of viral diseases like dengue fever, Zika and chikungunya. Aedes aegypti exhibits high morphological and behavioral variation, some of which is thought to be of epidemiological significance. Globally distributed domestic Ae. aegypti have often been grouped into (i the very pale variety queenslandensis and (ii the type form. Because the two color forms co-occur across most of their range, there is interest in understanding how freely they interbreed. This knowledge is particularly important for control strategies that rely on mating compatibilities between the release and target mosquitoes, such as Wolbachia releases and SIT. To address this question, we analyzed nuclear and mitochondrial genome-wide variation in the co-occurring pale and type Ae. aegypti from northern Queensland (Australia and Singapore.We typed 74 individuals at a 1170 bp-long mitochondrial sequence and at 16,569 nuclear SNPs using a customized double-digest RAD sequencing. 11/29 genotyped individuals from Singapore and 11/45 from Queensland were identified as var. queenslandensis based on the diagnostic scaling patterns. We found 24 different mitochondrial haplotypes, seven of which were shared between the two forms. Multivariate genetic clustering based on nuclear SNPs corresponded to individuals' geographic location, not their color. Several family groups consisted of both forms and three queenslandensis individuals were Wolbachia infected, indicating previous breeding with the type form which has been used to introduce Wolbachia into Ae. aegypti populations.Aedes aegypti queenslandensis are genomically indistinguishable from the type form, which points to these forms freely interbreeding at least in Australia and Singapore. Based on our findings, it is unlikely that the presence of very pale Ae. aegypti will affect the success of Aedes control programs based on Wolbachia-infected, sterile or RIDL mosquitoes.

The mosquitoAedes aegypti (L.) is a major vector of viral diseases like dengue fever, Zika and chikungunya. Aedes aegypti exhibits high morphological and behavioral variation, some of which is thought to be of epidemiological significance. Globally distributed domestic Ae. aegypti have often been grouped into (i) the very pale variety queenslandensis and (ii) the type form. Because the two color forms co-occur across most of their range, there is interest in understanding how freely they interbreed. This knowledge is particularly important for control strategies that rely on mating compatibilities between the release and target mosquitoes, such as Wolbachia releases and SIT. To address this question, we analyzed nuclear and mitochondrial genome-wide variation in the co-occurring pale and type Ae. aegypti from northern Queensland (Australia) and Singapore. We typed 74 individuals at a 1170 bp-long mitochondrial sequence and at 16,569 nuclear SNPs using a customized double-digest RAD sequencing. 11/29 genotyped individuals from Singapore and 11/45 from Queensland were identified as var. queenslandensis based on the diagnostic scaling patterns. We found 24 different mitochondrial haplotypes, seven of which were shared between the two forms. Multivariate genetic clustering based on nuclear SNPs corresponded to individuals' geographic location, not their color. Several family groups consisted of both forms and three queenslandensis individuals were Wolbachia infected, indicating previous breeding with the type form which has been used to introduce Wolbachia into Ae. aegypti populations. Aedes aegypti queenslandensis are genomically indistinguishable from the type form, which points to these forms freely interbreeding at least in Australia and Singapore. Based on our findings, it is unlikely that the presence of very pale Ae. aegypti will affect the success of Aedes control programs based on Wolbachia-infected, sterile or RIDL mosquitoes.

Full Text Available Zika virus (ZIKV, a flavivirus transmitted primarily by Aedes aegypti, has recently spread globally in an unprecedented fashion, yet we have a poor understanding of host-microbe interactions in this system. To gain insights into the interplay between ZIKV and the mosquito, we sequenced the small RNA profiles in ZIKV-infected and non-infected Ae. aegypti mosquitoes at 2, 7 and 14 days post-infection. ZIKA induced an RNAi response in the mosquito with virus-derived short interfering RNAs and PIWI-interacting RNAs dramatically increased in abundance post-infection. Further, we found 17 host microRNAs (miRNAs that were modulated by ZIKV infection at all time points. Strikingly, many of these regulated miRNAs have been reported to have their expression altered by dengue and West Nile viruses, while the response was divergent from that induced by the alphavirus Chikungunya virus in mosquitoes. This suggests that conserved miRNA responses occur within mosquitoes in response to flavivirus infection. This study expands our understanding of ZIKV-vector interactions and provides potential avenues to be further investigated to target ZIKV in the mosquito host.

Dengue fever is the most common mosquito transmitted viral infection afflicting humans, estimated to generate around 390 million infections each year in over 100 countries. The introduction of the endosymbiotic bacterium Wolbachia into Aedes aegypti mosquitoes has the potential to greatly reduce the public health burden of the disease. This approach requires extensive PCR (Polymerase Chain Reaction) testing of the Wolbachia-infection status of mosquitoes in areas where Wolbachia-A. aegypti are released. Here we report the first example of small organism mid-infrared spectroscopy where we have applied Attenuated Total Reflection Fourier Transform Infrared (ATR-FTIR) spectroscopy and multivariate modelling methods to determine sex, age and the presence of Wolbachia (wMel strain) in laboratory mosquitoes and sex and age in field mosquitoes. The prediction errors using Partial Least Squares Discriminant Analysis (PLS-DA) discrimination models for laboratory studies on independent test sets ranged from 0 to 3% for age & sex grading, and 3 to 5% for Wolbachia infection diagnosis using dry mosquito abdomens while field study results using an Artificial neural network yielded a 10 % error. The application of FTIR analysis is inexpensive, easy to use, portable, and shows significant potential to replace the reliance on more expensive and laborious PCR assays.

Full Text Available The chikungunya virus (CHIKV is an emerging pathogen with widespread distribution in regions of Africa, India, and Asia that threatens to spread into temperate climates with the introduction of its major vector, Aedes albopictus. CHIKV causes a disease frequently misdiagnosed as dengue fever, with potentially life-threatening symptoms that can result in a longer-term debilitating arthritis. The increasing risk of spread from endemic regions via human travel and commerce and the current absence of a vaccine put a significant proportion of the world population at risk for this disease. In this study we designed and tested hammerhead ribozymes (hRzs targeting CHIKV structural protein genes of the RNA genome as potential antivirals both at the cellular and in vivo level. We employed the CHIKV strain 181/25, which exhibits similar infectivity rates in both Vero cell cultures and mosquitoes. Virus suppression assay performed on transformed Vero cell clones of all seven hRzs demonstrated that all are effective at inhibiting CHIKV in Vero cells, with hRz #9 and #14 being the most effective. piggyBac transformation vectors were constructed using the Ae. aegypti t-RNAval Pol III promoted hRz #9 and #14 effector genes to establish a total of nine unique transgenic Higgs White Eye (HWE Ae. aegypti lines. Following confirmation of transgene expression by real-time polymerase chain reaction (RT-PCR, comparative TCID50-IFA analysis, in situ Immuno-fluorescent Assays (IFA and analysis of salivary CHIKV titers demonstrated effective suppression of virus replication at 7 dpi in heterozygous females of each of these transgenic lines compared with control HWE mosquitoes. This report provides a proof that appropriately engineered hRzs are powerful antiviral effector genes suitable for population replacement strategies

The chikungunya virus (CHIKV) is an emerging pathogen with widespread distribution in regions of Africa, India, and Asia that threatens to spread into temperate climates with the introduction of its major vector, Aedes albopictus. CHIKV causes a disease frequently misdiagnosed as dengue fever, with potentially life-threatening symptoms that can result in a longer-term debilitating arthritis. The increasing risk of spread from endemic regions via human travel and commerce and the current absence of a vaccine put a significant proportion of the world population at risk for this disease. In this study we designed and tested hammerhead ribozymes (hRzs) targeting CHIKV structural protein genes of the RNA genome as potential antivirals both at the cellular and in vivo level. We employed the CHIKV strain 181/25, which exhibits similar infectivity rates in both Vero cell cultures and mosquitoes. Virus suppression assay performed on transformed Vero cell clones of all seven hRzs demonstrated that all are effective at inhibiting CHIKV in Vero cells, with hRz #9 and #14 being the most effective. piggyBac transformation vectors were constructed using the Ae. aegypti t-RNAval Pol III promoted hRz #9 and #14 effector genes to establish a total of nine unique transgenic Higgs White Eye (HWE) Ae. aegypti lines. Following confirmation of transgene expression by real-time polymerase chain reaction (RT-PCR), comparative TCID50-IFA analysis, in situ Immuno-fluorescent Assays (IFA) and analysis of salivary CHIKV titers demonstrated effective suppression of virus replication at 7 dpi in heterozygous females of each of these transgenic lines compared with control HWE mosquitoes. This report provides a proof that appropriately engineered hRzs are powerful antiviral effector genes suitable for population replacement strategies PMID:27294950

Background The Asian tiger mosquito, Aedes albopictus is currently an important vector for dengue, chikungunya and Zika virus, and its role in transmission of arthropod-borne viruses (arboviruses) may increase in the future due to its ability to colonize temperate regions. In contrast to Aedes aegypti, the dominant vector of dengue, chikungunya and Zika virus, genetic responses of Ae. albopictus upon infection with an arbovirus are not well characterized. Here we present a study of the changes in transcript expression in Ae. albopictus exposed to dengue virus serotype 2 via feeding on an artificial bloodmeal. Methodology/Principal findings We isolated midguts and midgut-free carcasses of Ae. albopictus fed on bloodmeals containing dengue virus as well as controls fed on virus-free control meals at day 1 and day 5 post-feeding. We confirmed infection of midguts from mosquitoes sampled on day 5 post-feeding via RT-PCR. RNAseq analysis revealed dynamic modulation of the expression of several putative immunity and dengue virus-responsive genes, some of whose expression was verified by qRT-PCR. For example, a serine protease gene was up-regulated in the midgut at 1 day post infection, which may potentially enhance mosquito susceptibility to dengue infection, while 14 leucine-rich repeat genes, previously shown to be involved in mosquito antiviral defenses, were down-regulated in the carcass at 5 days post infection. The number of significantly modulated genes decreased over time in midguts and increased in carcasses. Conclusion/Significance Dengue virus exposure results in the modulation of genes in a time- and site-specific manner. Previous literature on the interaction between mosquitoes and mosquito-borne pathogens suggests that most of the changes that occurred in Ae. albopictus exposed to DENV would favor virus infection. Many genes identified in this study warrant further characterization to understand their role in viral manipulation of and antiviral response of

Mosquitoes are blood-feeding insects serving as the most important vectors for spreading human pathogens and parasites. Dengue is a viral disease mainly vectored through the bite of Aedesmosquitoes. Its transmission has recently increased in urban and semi-urban areas of tropical and subtropical regions worldwide, becoming a major international public health concern. There is no specific treatment for dengue. Its prevention and control solely depend on effective vector control measures. Mangrove plants have been used in Indian traditional medicine for a wide array of purposes. In this research, we proposed a method for biosynthesis of antiviral and mosquitocidal silver nanoparticles (AgNP) using the aqueous extract of Bruguiera cylindrica leaves. AgNP were characterized using a variety of biophysical analyses, including UV-visible spectrophotometry, Fourier-transform infrared spectroscopy, scanning electron microscopy, and energy-dispersive X-ray spectroscopy. Bruguiera cilyndrica aqueous extract and green-synthesized AgNP were tested against the primary dengue vector Aedes aegypti. AgNP were the most effective. LC50 values ranged from 8.93 ppm (larva I) to 30.69 ppm (pupa). In vitro experiments showed that 30 μg/ml of AgNP significantly inhibited the production of dengue viral envelope (E) protein in vero cells and downregulated the expression of dengue viral E gene. Concerning nontarget effects, we observed that the predation efficiency of Carassius auratus against A. aegypti was not affected by exposure at sublethal doses of AgNP. Predation in the control was 71.81 % (larva II) and 50.43 % (larva III), while in an AgNP-treated environment, predation was boosted to 90.25 and 76.81 %, respectively. Overall, this study highlights the concrete potential of green-synthesized AgNP in the fight against dengue virus. Furthermore, B. cylindrica-synthesized AgNP can be employed at low doses to reduce larval and pupal population of A. aegypti, without detrimental

We report computational simulations for the evolution of the population of the dengue vector, Aedes aegypti mosquitoes. The results suggest that controlling the mosquito population, on the basis of intraspecific competition at the larval stage, can be an efficient mechanism for controlling the spread of the epidemic. The results also show the presence of a kind of genetic evolution in vector population, which results mainly in increasing the average lifespan of individuals in adulthood.

Full Text Available BACKGROUND: Hematophagy poses a challenge to blood-feeding organisms since products of blood digestion can exert cellular deleterious effects. Mitochondria perform multiple roles in cell biology acting as the site of aerobic energy-transducing pathways, and also an important source of reactive oxygen species (ROS, modulating redox metabolism. Therefore, regulation of mitochondrial function should be relevant for hematophagous arthropods. Here, we investigated the effects of blood-feeding on flight muscle (FM mitochondria from the mosquitoAedes aegypti, a vector of dengue and yellow fever. METHODOLOGY/PRINCIPAL FINDINGS: Blood-feeding caused a reversible reduction in mitochondrial oxygen consumption, an event that was parallel to blood digestion. These changes were most intense at 24 h after blood meal (ABM, the peak of blood digestion, when oxygen consumption was inhibited by 68%. Cytochromes c and a+a(3 levels and cytochrome c oxidase activity of the electron transport chain were all reduced at 24 h ABM. Ultrastructural and molecular analyses of FM revealed that mitochondria fuse upon blood meal, a condition related to reduced ROS generation. Consistently, BF induced a reversible decrease in mitochondrial H(2O(2 formation during blood digestion, reaching their lowest values at 24 h ABM where a reduction of 51% was observed. CONCLUSION: Blood-feeding triggers functional and structural changes in hematophagous insect mitochondria, which may represent an important adaptation to blood feeding.

Full Text Available Enantiomers differ only in the left or right handedness (chirality of their orientations and exhibit identical chemical and physical properties. In chemical communication systems, enantiomers can be differentially active at the physiological and behavioral levels. Only recently were enantioselective odorant receptors demonstrated in mammals while their existence in insects has remained hypothetical. Using the two-microelectrode voltage clamp of Xenopus oocytes, we show that the yellow fever mosquito, Aedes aegypti, odorant receptor 8 (AaOR8 acts as a chiral selective receptor for the (R-(--enantiomer of 1-octen-3-ol, which in the presence of other kairomones is an attractant used by blood-sucking insects to locate their hosts. In addition to steric constraints, chain length and degree of unsaturation play important roles in this recognition process. This is the first characterization of an enantioselective odorant receptor in insects and the results demonstrate that an OR alone, without helper proteins, can account for chiral specificity exhibited by olfactory sensory neurons (OSNs.

Injury and infection priming has been observed in several insect groups, reported as host immune protection against contact with a pathogen caused by a previous infection with the same. However, the specific response against a pathogen has not been demonstrated in all insect species. Investigating the specific priming response in insects is important because their immune strategies probably reflect particular selective pressures exerted by different pathogens. Here, we determined whether previous infection of Aedes aegypti would enhance survival and/or lead to greater and specific AMP expression after a second exposure to the same or a distinct bacterium. Mosquitoes previously immunized with a low dose of Escherichia coli, but not Staphylococcus aureus, showed increased survival. Although the host protection herein demonstrated was not specific, each bacterium elicited differential AMP expression. These results can be explained by the susceptible-primed-infected (SPI) epidemiological model, which poses that in the evolution of memory-like responses (priming), a pivotal role is played by pathogen virulence, associated host damage, and the host capacity of pathogen recognition. PMID:27446016

Full Text Available Background & objectives: Although having immense clinical relevance, yet only a few studies have been targeted to understand the chikungunya virus (CHIKV susceptibility and growth in Aedes aegypti populations from India. This study was undertaken to investigate CHIKV susceptibility and growth kinetics in Ae. aegypti along with genetic heterogeneity of Ae. aegypti populations. Methods: Dose dependent CHIKV susceptibility and growth kinetic studies for three CHIKV strains reported from India were carried out in Ae. aegypti mosquito populations. The phenotypic variation and genetic heterogeneity in five Ae. aegypti populations were investigated using multivariate morphometrics and allozyme variation studies. Results: The dissemination and growth kinetics studies of the three CHIKV strains showed no selective advantage for a particular strain of CHIKV in Ae. aegypti. At 100 per cent infection rate, five geographic Ae. aegypti populations showed differences in dissemination to three CHIKV strains. Morphometric studies revealed phenotypic variation in all the studied populations. The allelic frequencies, F statistics, and Nei′s genetic identity values showed that genetic differences between the populations were small, but significant. Interpretation & conclusions: The results obtained in this study suggest that genetic background of the vector strongly influences the CHIKV susceptibility in Ae. aegypti.

Aedes aegypti (L.) mosquitoes preferentially oviposit in natural and artificial receptacles where their eggs are able to withstand drying as water levels fluctuate. Desiccation-resistant eggs also increase the potential for establishment in non-native habitats while providing logistical impediments to control programs. Viability and mean survival times of eggs stored under three dryness conditions for up to 367 days were investigated among three field-derived colonies of Australian Ae. aegypti to understand variation in desiccation survival. Further investigations compared egg survival between an established colony and its wild counterpart. Our results confirmed that Ae. aegypti eggs can withstand desiccation for extended periods of time with approximately 2-15% egg viability recorded after one year and viability remaining above 88% under all conditions through 56 days. Intraspecific variations in egg survival times were recorded, suggesting local adaptation while each of the colonies demonstrated a consistent preference for higher humidity. Egg volume varied between the populations, suggesting a relationship between egg volume and survival time, with the marginally larger eggs (Charters Towers and Innisfail) having greater desiccation resistance over the range of conditions. The strong survivorship of Charters Towers eggs in dry, warm conditions demonstrates the adaptive significance of a desiccation-resistant egg.

Background Hematophagy poses a challenge to blood-feeding organisms since products of blood digestion can exert cellular deleterious effects. Mitochondria perform multiple roles in cell biology acting as the site of aerobic energy-transducing pathways, and also an important source of reactive oxygen species (ROS), modulating redox metabolism. Therefore, regulation of mitochondrial function should be relevant for hematophagous arthropods. Here, we investigated the effects of blood-feeding on flight muscle (FM) mitochondria from the mosquitoAedes aegypti, a vector of dengue and yellow fever. Methodology/Principal Findings Blood-feeding caused a reversible reduction in mitochondrial oxygen consumption, an event that was parallel to blood digestion. These changes were most intense at 24 h after blood meal (ABM), the peak of blood digestion, when oxygen consumption was inhibited by 68%. Cytochromes c and a+a3 levels and cytochrome c oxidase activity of the electron transport chain were all reduced at 24 h ABM. Ultrastructural and molecular analyses of FM revealed that mitochondria fuse upon blood meal, a condition related to reduced ROS generation. Consistently, BF induced a reversible decrease in mitochondrial H2O2 formation during blood digestion, reaching their lowest values at 24 h ABM where a reduction of 51% was observed. Conclusion Blood-feeding triggers functional and structural changes in hematophagous insect mitochondria, which may represent an important adaptation to blood feeding. PMID:19924237

We have initiated efforts to determine the molecular basis for the M(D) meiotic drive system in the mosquito, Aedes aegypti. The effect of the M(D) gene is a highly male-biased sex ratio, but varies depending on the frequency and sensitivity of a susceptible responder m(s) allele. The M(D) system has potential as a mechanism for driving trangenes for pathogen resistance into natural Ae. aegypti populations. Because all previously existing laboratory strains carrying the M(D) gene have been lost, we have selected for a new strain, T37, that carries a strong driver. Matings between T37 males and drive-susceptible m(s) females result in progeny with highly biased sex ratios, wherein only approximately 14.7% females are produced. We discuss the potential for identifying M(D) candidate genes based on comparisons with the well-described Drosophila melanogaster segregation distorter (SD) meiotic drive system and considerations for release of transgenic Ae. aegypti into natural populations where M(D) and insensitive m3 alleles are likely segregating.

Full Text Available The mosquitoAedes aegypti is a potent vector of the chikungunya, yellow fever, and dengue viruses, responsible for hundreds of millions of infections and over 50,000 human deaths per year. Mutagenesis in Ae. aegypti has been established with TALENs, ZFNs, and homing endonucleases, which require the engineering of DNA-binding protein domains to provide genomic target sequence specificity. Here, we describe the use of the CRISPR-Cas9 system to generate site-specific mutations in Ae. aegypti. This system relies on RNA-DNA base-pairing to generate targeting specificity, resulting in efficient and flexible genome-editing reagents. We investigate the efficiency of injection mix compositions, demonstrate the ability of CRISPR-Cas9 to generate different types of mutations via disparate repair mechanisms, and report stable germline mutations in several genomic loci. This work offers a detailed exploration into the use of CRISPR-Cas9 in Ae. aegypti that should be applicable to non-model organisms previously out of reach of genetic modification.

Objective:To evaluate the effects of deltamethrin against field-collected adults of Aedes aegypti L. (Ae. aegypti). Methods: The adults were selected with 0.025%deltamethrin for 40 successive generations. The selected adults were tested with 4%DDT and the emerging larvae were tested with various insecticides to study the cross-resistance spectrum. The knockdown and irritability studies were carried out in adult mosquitoes to investigate their behavioural response to deltamethrin. Results:Forty generations of selection with deltamethrin resulted in only 3.8-fold resistance in the adults of Ae. aegypti. The adults of parent (PS) and selected strains (DAS) exhibited only 0.8-fold cross resistance to 4%DDT. The larvae emerging from the PS and DAS strains did not develop appreciable levels of resistance to various insecticides tested. The knockdown studies revealed KT50 of 14.4 min in PS adults with no signs of recovery even after 24 h. The DAS strains could develop only 1.2 to 1.3-fold knockdown resistance (KDR). The knockdown response of DDT was though 5-6 times slower than deltamethrin but the continued response in deltamethrin-selected adults caused only 1.2-fold KDR. The PS and DAS strains exhibited significant irritability response towards deltamethrin and DDT. The DAS strains showed 5-6 fold increased irritability to deltamethrin as compared to the PS strain. Conclusions:The above results suggest the prolonged effective use of deltamethrin against Ae. aegypti as an adulticide.

The currently spreading arbovirus epidemic is having a severe impact on human health worldwide. The two most common flaviviruses, dengue virus (DENV) and Zika virus (ZIKV), are transmitted through the same viral vector, Aedes spp. mosquitoes. Since the discovery of DENV in 1943, this virus has been reported to cause around 390 million human infections per year, approximately 500,000 of which require hospitalization and over 20,000 of which are lethal. The present DENV epidemic is primarily concentrated in Southeast Asia. ZIKV, which was discovered in 1952, is another important arthropod-borne flavivirus. The neurotropic role of ZIKV has been reported in infected newborns with microcephaly and in adults with Guillain-Barre syndrome. Despite DENV and ZIKV sharing the same viral vector, their complex pathogenic natures are poorly understood, and the infections they cause do not have specific treatments or effective vaccines. Therefore, this review will describe what is currently known about the clinical characteristics, pathogenesis mechanisms, and transmission of these two viruses. Better understanding of the interrelationships between DENV and ZIKV will provide a useful perspective for developing an effective strategy for controlling both viruses in the future.

The Asian tiger mosquitoAedes albopictus is currently one of the most threatening invasive species in the world. Native to Southeast Asia, the species has spread throughout the world in the past 30 years and is now present in every continent but Antarctica. Because it was the main vector of recent Dengue and Chikungunya outbreaks, and because of its competency for numerous other viruses and pathogens such as the Zika virus, A. albopictus stands out as a model species for invasive diseases vector studies. A synthesis of the current knowledge about the genetic diversity of A. albopictus is needed, knowing the interplays between the vector, the pathogens, the environment and their epidemiological consequences. Such resources are also valuable for assessing the role of genetic diversity in the invasive success. We review here the large but sometimes dispersed literature about the population genetics of A. albopictus. We first debate about the experimental design of these studies and present an up-to-date assessment of the available molecular markers. We then summarize the main genetic characteristics of natural populations and synthesize the available data regarding the worldwide structuring of the vector. Finally, we pinpoint the gaps that remain to be addressed and suggest possible research directions.

Background A five-year citywide control program based on regular application of temephos significantly reduced Aedes aegypti larval indices but failed to maintain them below target levels in Clorinda, northern Argentina. Incomplete surveillance coverage and reduced residuality of temephos were held as the main putative causes limiting effectiveness of control actions. Methodology The duration of temephos residual effects in household-owned water-holding tanks (the most productive container type and main target for control) was estimated prospectively in two trials. Temephos was applied using spoons or inside perforated small zip-lock bags. Water samples from the study tanks (including positive and negative controls) were collected weekly and subjected to larval mortality bioassays. Water turnover was estimated quantitatively by adding sodium chloride to the study tanks and measuring its dilution 48 hs later. Principal Findings The median duration of residual effects of temephos applied using spoons (2.4 weeks) was significantly lower than with zip-lock bags (3.4 weeks), and widely heterogeneous between tanks. Generalized estimating equations models showed that bioassay larval mortality was strongly affected by water type and type of temephos application depending on water type. Water type and water turnover were highly significantly associated. Tanks filled with piped water had high turnover rates and short-lasting residual effects, whereas tanks filled with rain water showed the opposite pattern. On average, larval infestations reappeared nine weeks post-treatment and seven weeks after estimated loss of residuality. Conclusions Temephos residuality in the field was much shorter and more variable than expected. The main factor limiting temephos residuality was fast water turnover, caused by householders' practice of refilling tanks overnight to counteract the intermittence of the local water supply. Limited field residuality of temephos accounts in part for the

Within the last 10 years Zika virus (ZIKV) has caused unprecedented epidemics of human disease in the nations and territories of the western Pacific and South America, and continues to escalate in both endemic and non-endemic regions. We evaluated the vector competence of Australian mosquitoes for ZIKV to assess their potential role in virus transmission. Mosquitoes were exposed to infectious blood meals containing the prototype African ZIKV strain. After 14 days incubation at 28°C and high relative humidity, infection, dissemination and transmission rates were assessed. Infection in Culex annulirostris and Cx. sitiens could not be detected. 8% of Cx. quinquefasciatus were infected, but the virus did not disseminate in this species. Despite having infection rates > 50%, Aedes notoscriptus and Ae. vigilax did not transmit ZIKV. In contrast, Ae. aegypti had infection and transmission rates of 57% and 27%, respectively. In susceptibility trials, the virus dose required to infect 50% (ID50) of Ae. aegypti was106.4 tissue culture infectious dose50 (TCID50)/mL. Additionally, a threshold viral load within the mosquito of at least 105.1 TCID50 equivalents/mL had to be reached before virus transmission occurred. We confirmed Ae. aegypti to be the most likely mosquito vector of ZIKV in Australia, although the restricted distribution of this species will limit the receptive zone to northern Queensland where this species occurs. Importantly, the role in ZIKV transmission of Culex and other Aedes spp. tested will be negligible. Despite being the implicated vector, the relatively high ID50 and need for a high titer disseminated infection in Ae. aegypti suggest that high mosquito population densities will be required to facilitate epidemic ZIKV transmission among the currently immunologically naïve human population in Australia.

Full Text Available Within the last 10 years Zika virus (ZIKV has caused unprecedented epidemics of human disease in the nations and territories of the western Pacific and South America, and continues to escalate in both endemic and non-endemic regions. We evaluated the vector competence of Australian mosquitoes for ZIKV to assess their potential role in virus transmission.Mosquitoes were exposed to infectious blood meals containing the prototype African ZIKV strain. After 14 days incubation at 28°C and high relative humidity, infection, dissemination and transmission rates were assessed. Infection in Culex annulirostris and Cx. sitiens could not be detected. 8% of Cx. quinquefasciatus were infected, but the virus did not disseminate in this species. Despite having infection rates > 50%, Aedes notoscriptus and Ae. vigilax did not transmit ZIKV. In contrast, Ae. aegypti had infection and transmission rates of 57% and 27%, respectively. In susceptibility trials, the virus dose required to infect 50% (ID50 of Ae. aegypti was106.4 tissue culture infectious dose50 (TCID50/mL. Additionally, a threshold viral load within the mosquito of at least 105.1 TCID50 equivalents/mL had to be reached before virus transmission occurred.We confirmed Ae. aegypti to be the most likely mosquito vector of ZIKV in Australia, although the restricted distribution of this species will limit the receptive zone to northern Queensland where this species occurs. Importantly, the role in ZIKV transmission of Culex and other Aedes spp. tested will be negligible. Despite being the implicated vector, the relatively high ID50 and need for a high titer disseminated infection in Ae. aegypti suggest that high mosquito population densities will be required to facilitate epidemic ZIKV transmission among the currently immunologically naïve human population in Australia.

Background Within the last 10 years Zika virus (ZIKV) has caused unprecedented epidemics of human disease in the nations and territories of the western Pacific and South America, and continues to escalate in both endemic and non-endemic regions. We evaluated the vector competence of Australian mosquitoes for ZIKV to assess their potential role in virus transmission. Methodology/Principal Findings Mosquitoes were exposed to infectious blood meals containing the prototype African ZIKV strain. After 14 days incubation at 28°C and high relative humidity, infection, dissemination and transmission rates were assessed. Infection in Culex annulirostris and Cx. sitiens could not be detected. 8% of Cx. quinquefasciatus were infected, but the virus did not disseminate in this species. Despite having infection rates > 50%, Aedes notoscriptus and Ae. vigilax did not transmit ZIKV. In contrast, Ae. aegypti had infection and transmission rates of 57% and 27%, respectively. In susceptibility trials, the virus dose required to infect 50% (ID50) of Ae. aegypti was106.4 tissue culture infectious dose50 (TCID50)/mL. Additionally, a threshold viral load within the mosquito of at least 105.1 TCID50 equivalents/mL had to be reached before virus transmission occurred. Conclusions/Significance We confirmed Ae. aegypti to be the most likely mosquito vector of ZIKV in Australia, although the restricted distribution of this species will limit the receptive zone to northern Queensland where this species occurs. Importantly, the role in ZIKV transmission of Culex and other Aedes spp. tested will be negligible. Despite being the implicated vector, the relatively high ID50 and need for a high titer disseminated infection in Ae. aegypti suggest that high mosquito population densities will be required to facilitate epidemic ZIKV transmission among the currently immunologically naïve human population in Australia. PMID:27643685

After female mosquitoes ingest blood from vertebrate hosts, exopeptidases and endopeptidases are required for digesting blood proteins in the midgut into amino acids, which female mosquitoes use to build yolk proteins. These proteases are not always present in the midgut, and their diverse expression patterns suggest that production of these enzymes is highly regulated in order to meet specific physiological demands at various stages. Here we report identification of a serine-type protease, JHA15, in the yellow fever mosquitoAedes aegypti. This protein shares high sequence homology with chymotrypsins, and indeed exhibits specific chymotrypsin enzymatic activity. The JHA15 gene is expressed primarily in the midgut of adult female mosquitoes. Our results indicate that its transcription is activated by juvenile hormone in the newly emerged female adults. Although its mRNA profile is similar to that of the early trypsin gene, we found that JHA15 proteins were readily detected in the midgut epithelium cells of both non-blood-fed and blood-fed mosquitoes. Analysis of polysomal RNA further substantiated that synthesis of JHA15 occurs before and shortly after blood feeding. Knocking down expression of JHA15 resulted in no evident phenotypic changes, implying that functional redundancy exists among those proteolytic enzymes.

Mosquitoes are rapidly developing resistance to insecticides that millions of people relied on to protect themselves from the diseases they carry, thereby creating a need to develop new insecticides. Clausena anisata is used traditionally as an insect repellent by various communities in Africa and Asia. For this study, the repellency and adulticidal activities of leaf extracts and compounds isolated from this plant species were evaluated against the yellow fever mosquito, Aedes aegypti. In the topical application assays, using total bites as an indicator, repellency was dose dependent, with the acetone crude extract (15 %) having 93 % repellence and the hexane fraction (7.5 %) 67 % repellence after 3 h. Fractionation resulted in a loss of total repellence. As mosquito-net treating agents, the acetone and hexane extracts of C. anisata, both at 15 %, had average repellences of 46.89 ± 2.95 and 50.13 ± 2.02 %, respectively, 3 h after exposure. The C. anisata acetone extract and its hexane fraction caused mosquito knockdown and eventually death when nebulised into the testing chamber, with an EC50 of 78.9 mg/ml (7.89 %) and 71.6 mg/ml (7.16 %) in the first 15 min after spraying. C. anisata leaf extracts have potential to be included in protection products against mosquitoes due to the repellent and cidal compounds contained therein.

A large proportion of mosquitolarval habitats in urban and rural communities in sub-Saharan Africa are man-made. Therefore, community-based larval source management (LSM) could make a significant contribution to malaria control in an integrated vector management approach. Here we implemented an exp

Mosquito-borne diseases with an economic impact create loss in commercial and labor outputs, particularly in countries with tropical and subtropical climates. Mosquito control is facing a threat because of the emergence of resistance to synthetic insecticides. Extracts from plants may be alternative sources of mosquito control agents because they constitute a rich source of bioactive compounds that are biodegradable into nontoxic products and potentially suitable for use to control mosquitoes. Insecticides of botanical origin may serve as suitable alternative biocontrol techniques in the future. In view of the recently increased interest in developing plant origin insecticides as an alternative to chemical insecticide, this study was undertaken to assess the larvicidal and ovicidal potential of the crude hexane, benzene, chloroform, ethyl acetate, and methanol solvent extracts from the medicinal plant Delonix elata against the medically important mosquito vectors, Anopheles stephensi and Aedes aegypti (Diptera: Culicidae). The larval mortality was observed after 24 h of exposure. All extracts showed moderate larvicidal effects; however, the highest larval mortality was found in methanol extract of leaf of D. elata against the larvae of A. stephensi and A. aegypti with the LC(50) and LC(90) values being 93.59 and 111.83, and 163.69 and 202.77 ppm, respectively. Compared to leaf extracts, seeds have low potency against two mosquitoes with the LC(50) and LC(90) values being 115.28 and 139.04, and 225.07 and 273.03 ppm, respectively. The mean percent hatchability of the eggs was observed after 48 h post-treatment. The percent hatchability was inversely proportional to the concentration of extract and directly proportional to the eggs. All the five solvent extracts showed moderate ovicidal activity; however, the methanol extract showed the highest ovicidal activity. One hundred percent mortality was observed at 300 ppm for leaf methanol extract and 500 ppm for seed

Objective: To determine the effects of environmental characteristics of larval habitats on distribution and abundance of anopheline mosquitoes in Bashagard county, a malarious area in southeast of Iran. Methods: Larvae were collected monthly using the standard dipping method and identified using a morphological-based key. Environmental characteristics of the larval habitats were recorded. Water samples were taken from habitats during larval collection for physico-chemical characterization. Statistical analyses were performed. Results: In total 5150 anopheline larvae from 36 larval habitats were collected and identified. They comprised of six species: Anopheles culicifacies (29.36%), Anopheles moghulensis (25.20%),Anopheles dthali stephensi (5.01%). (18.02%), Anopheles superpictus (17.24%), Anopheles turkhudi (5.17%) and Anopheles The most common larval habitats were natural and clear water bodies such as riverbeds with sandy substrates and still water. Furthermore, the anopheline larvae were abundant in permanent and full sunlight habitats without vegetation and algae. Larval density was positively correlated with water temperature. Chemical characteristics including conductivity, total alkalinity, sulphate and chloride had significant effects on distribution and abundance of anopheline species.Conclusions:The result of this study indicates a correlation between some environmental characteristics and anopheline larvae abundance which can be considered for effective planning and implementing malaria elimination program in Iran.

The orphan nuclear receptor HR3 is essential for developmental switches during insect development and metamorphosis regulated by 20-hydroxyecdysone (20E). Reproduction of female mosquitoes of the major vector of Dengue fever, Aedes aegypti, is cyclic because of its dependence on blood feeding. 20E is an important hormone regulating vitellogenic events in this mosquito; however, any role for HR3 in 20E-driven reproductive events has not been known. Using RNA interference (RNAi) approach, we demonstrated that Aedes HR3 plays a critical role in a timely termination of expression of the vitellogenin (Vg) gene encoding the major yolk protein precursor. It is also important for downregulation of the Target-of-Rapamycin pathway and activation of programmed autophagy in the Aedes fat body at the end of vitellogenesis. HR3 is critical in activating betaFTZ-F1, EcRB and USPA, the expressions of which are highly elevated at the end of vitellogenesis. RNAi depletion of HR3 (iHR3) prior to the first gonadotrophic cycle affects a normal progression of the second gonadotrophic cycle. Most of ovaries 24 h post second blood meal from iHR3 females in the second cycle were small with follicles that were only slightly different in length from of those of resting stage. In addition, these iHR3 females laid a significantly reduced number of eggs per mosquito as compared to those of iMal and the wild type. Our results indicate an important role of HR3 in regulation of 20E-regulated developmental switches during reproductive cycles of A. aegypti females.

Full Text Available The orphan nuclear receptor HR3 is essential for developmental switches during insect development and metamorphosis regulated by 20-hydroxyecdysone (20E. Reproduction of female mosquitoes of the major vector of Dengue fever, Aedes aegypti, is cyclic because of its dependence on blood feeding. 20E is an important hormone regulating vitellogenic events in this mosquito; however, any role for HR3 in 20E-driven reproductive events has not been known. Using RNA interference (RNAi approach, we demonstrated that Aedes HR3 plays a critical role in a timely termination of expression of the vitellogenin (Vg gene encoding the major yolk protein precursor. It is also important for downregulation of the Target-of-Rapamycin pathway and activation of programmed autophagy in the Aedes fat body at the end of vitellogenesis. HR3 is critical in activating betaFTZ-F1, EcRB and USPA, the expressions of which are highly elevated at the end of vitellogenesis. RNAi depletion of HR3 (iHR3 prior to the first gonadotrophic cycle affects a normal progression of the second gonadotrophic cycle. Most of ovaries 24 h post second blood meal from iHR3 females in the second cycle were small with follicles that were only slightly different in length from of those of resting stage. In addition, these iHR3 females laid a significantly reduced number of eggs per mosquito as compared to those of iMal and the wild type. Our results indicate an important role of HR3 in regulation of 20E-regulated developmental switches during reproductive cycles of A. aegypti females.

Objective: To identify the larvicidal activity of the volatile oil from Sesuvium portulacastrum (S.portulacastrum ) against Anopheles stephensi and Aedes aegypti. Methods: Volatile oil extract of S. portulacastrum was prepared in a graded series of concentration. The test for the larvicidal effect of volatile oil against mosquitos larvae was conducted in accordance with the WHO standard method. Batches of 25 early 4th instar larvae of two mosquitoes were transferred to 250 mL enamel bowl containing 199 mL of distilled water and 1 mL of plant extracts. Each experiment was conducted with triplicate with concurrent a control group.Results:Volatile oil extract of S. portulacastrum showed toxicity against 4th instar larvae of Aedes aegypti and Anopheles stephensi with equivalent LC50 value [(63±7.8) µL/mL, LCL-UCL=55.2-64.0] and LC90 value [(94.2±3.9) µL/mL)] in maximum activity with minimum concentration (200 µL/mL) of volatile oil and followed by maximum activity of 250 µL concentration showed LC50 value=(68.0±8.2) µL/mL, LCL-UCL=66.26-69.2 and LC50 value of (55.2±2.8) µL/mL, LCL-UCL=53.7-56.9, LC90=(95.2±1.25) µL/mL and followed by 250 µL of oil extract against 4th instar larvae of Aedes aegypti respectively.Conclusions:It is inferred from the present study that, the extracts from salt marsh mangrove plan of S. portulacastum are identified as a potential source of safe and efficacious mosquito control agents for the management of vector borne diseases of malaria and dengue.

The yellow fever mosquitoAedes aegypti is an important vector of viral diseases that impact global health. Insecticides are typically used to manage mosquito populations, but the evolution of insecticide resistance is limiting their effectiveness. Thus, identifying new molecular and physiological targets in mosquitoes is needed to facilitate insecticide discovery and development. Here we test the hypothesis that gap junctions are valid molecular and physiological targets for new insecticides. Gap junctions are intercellular channels that mediate direct communication between neighboring cells and consist of evolutionarily distinct proteins in vertebrate (connexins) and invertebrate (innexins) animals. We show that the injection of pharmacological inhibitors of gap junctions (i.e., carbenoxolone, meclofenamic acid, or mefloquine) into the hemolymph of adult female mosquitoes elicits dose-dependent toxic effects, with mefloquine showing the greatest potency. In contrast, when applied topically to the cuticle, carbenoxolone was the only inhibitor to exhibit full efficacy. In vivo urine excretion assays demonstrate that both carbenoxolone and mefloquine inhibit the diuretic output of adult female mosquitoes, suggesting inhibition of excretory functions as part of their mechanism of action. When added to the rearing water of 1st instar larvae, carbenoxolone and meclofenamic acid both elicit dose-dependent toxic effects, with meclofenamic acid showing the greatest potency. Injecting a double-stranded RNA cocktail against innexins into the hemolymph of adult female mosquitoes knock down whole-animal innexin mRNA expression and decreases survival of the mosquitoes. Taken together these data indicate that gap junctions may provide novel molecular and physiological targets for the development of insecticides.

Full Text Available The yellow fever mosquitoAedes aegypti is an important vector of viral diseases that impact global health. Insecticides are typically used to manage mosquito populations, but the evolution of insecticide resistance is limiting their effectiveness. Thus, identifying new molecular and physiological targets in mosquitoes is needed to facilitate insecticide discovery and development. Here we test the hypothesis that gap junctions are valid molecular and physiological targets for new insecticides. Gap junctions are intercellular channels that mediate direct communication between neighboring cells and consist of evolutionarily distinct proteins in vertebrate (connexins and invertebrate (innexins animals. We show that the injection of pharmacological inhibitors of gap junctions (i.e., carbenoxolone, meclofenamic acid, or mefloquine into the hemolymph of adult female mosquitoes elicits dose-dependent toxic effects, with mefloquine showing the greatest potency. In contrast, when applied topically to the cuticle, carbenoxolone was the only inhibitor to exhibit full efficacy. In vivo urine excretion assays demonstrate that both carbenoxolone and mefloquine inhibit the diuretic output of adult female mosquitoes, suggesting inhibition of excretory functions as part of their mechanism of action. When added to the rearing water of 1st instar larvae, carbenoxolone and meclofenamic acid both elicit dose-dependent toxic effects, with meclofenamic acid showing the greatest potency. Injecting a double-stranded RNA cocktail against innexins into the hemolymph of adult female mosquitoes knock down whole-animal innexin mRNA expression and decreases survival of the mosquitoes. Taken together these data indicate that gap junctions may provide novel molecular and physiological targets for the development of insecticides.

Acoustic larvicide devices are part of an emerging technology that provides a non-chemical and non-biological means to reduce larval populations of key medically important mosquito species such as Aedes aegypti in containers or catchments of water. These devices could benefit integrated vector manag...

Full Text Available A German traveler developed dengue fever in late August 2013, following a direct flight from Germany. Autochthonous dengue virus (DENV infection has not been reported in Japan. To evaluate the risk of autochthonous DENV transmission in Japan, the authors performed a retrospective search of the five areas visited by the German patient to determine the population density of dengue vector mosquito, Aedes albopictus. The annual mean temperature of each area was higher than 12 °C, which is considered suitable for the establishment of A. albopictus populations. Our retrospective search revealed the population density of A. albopictus to be high in the urban areas of Japan.

Background - Genetic modification of mosquitoes offers a promising strategy for the prevention and control of mosquito-borne diseases. For such a strategy to be effective, it is critically important that engineered strains are competitive enough to serve their intended function in population

Background - Genetic modification of mosquitoes offers a promising strategy for the prevention and control of mosquito-borne diseases. For such a strategy to be effective, it is critically important that engineered strains are competitive enough to serve their intended function in population replace

Full Text Available Mosquito vectors spread severe human diseases which lead to millions of deaths every year. Vector management is ultimately aimed to develop the health of every individual’s life by reducing the mosquito diversity. Control of vectors in growing counties is an important issue with various aspects. The advancement of green nanotechnology will attribute the solution for vector controlling policy. To identify the larvicidal property of silver nanoparticles (AgNPs using Belosynapsis kewensis (B. kewensis leaf extract against the Anopheles stephensi (A. stephensi and Aedes aegypti (A. aegypti in vitro study (LC50 and LC90 was analyzed. The synthesized AgNPs were characterized by UV–vis. (Ultraviolet–visible spectroscopy, FTIR (Fourier Transform Infra Red spectroscopy, TEM (Transmission Electron Microscopy, and XRD (X-ray Diffraction. Green AgNPs have a maximum absorption at 411 nm. The FTIR spectrum showed prominent peaks in (3863.55, 3759.02, 3361.01, 2926.81, 1575.12, 1388.16, 1034.79, 821.96, 717.07, 590.92, 534.32 and 472.42 cm−1 in the region of 4000–400 cm−1. The XRD peaks shown at 27.4°, 35.90°, 37.20°, 51.23° and 71.10° correspond to (311, (100, (101, (104 and (006 planes for face centered cubic (FCC. The TEM image showed the NPs with an average size of 24 ± 1.6 nm. In vitro larvicidal activity of AgNPs was used against the fourth instar of A. stephensi and A. aegypti. The LC50 and LC90 values of AgNPs showed to be effective against A. stephensi (LC50 = 78.4; LC90 = 144.7 ppm followed by A. aegypti (LC50 = 84.2; LC90 = 117.3 ppm. These results recommend that the green synthesized AgNPs have a potential to be used as a candidate for the control of A. stephensi and A. aegypti through eco-friendly and cost effective approach.

The hypothesis that size-selective predation and species-specific prey behaviours facilitate the coexistence between larvae of invasive Aedes albopictus (Skuse) and U.S.A.-native Ochlerotatus triseriatus (Say) was tested experimentally with the predator Corethrella appendiculata (Grabham).Larval behaviours associated with a higher risk of predation were identified, and prey behavioural responses were tested in either the physical presence of predators or in water containing predation cues. Larvae that thrashed on container bottoms had a higher risk of being captured by fourth instar C. appendiculata than did larvae resting on the water surface. Ochlerotatus triseriatus, but not A. albopictus, adopted low-risk behaviours in response to water-borne cues to predation. Both prey species reduced risky behaviours in the physical presence of the predator, but O. triseriatus showed a stronger response.The vulnerability of 2nd and 3rd instar prey to predation was compared, and behavioural responses were correlated with prey vulnerability. Second instars of both species were more vulnerable to predation by C. appendiculata than were 3rd instars, and the 3rd instar A. albopictus was more vulnerable than O. triseriatus of the same stage. All instars of O. triseriatus showed a similar reduction of risky behaviours in response to the presence of C. appendiculata despite 4th instar prey being relatively invulnerable to size-selective predation.Weaker predator avoidance, coupled with superior competitive ability, of invasive A. albopictus is likely to contribute to its coexistence with O. triseriatus in containers of the south-eastern U.S.A., where C. appendiculata can be abundant.

The coastal mosquitoAedes togoi occurs more or less continuously from subarctic to subtropic zones along the coasts of the Japanese islands and the East Asian mainland. It occurs also in tropical Southeast Asia and the North American Pacific coast, and the populations there are thought to have been introduced from Japan by ship. To test this hypothesis, the genetic divergence among geographic populations of A. togoi was studied using one mitochondrial and three nuclear gene sequences. We detected 71 mitochondrial haplotypes forming four lineages, with high nucleotide diversity around temperate Japan and declining towards peripheral ranges. The major lineage (L1) comprised 57 haplotypes from temperate and subarctic zones in Japan and Southeast Asia including southern China and Taiwan. Two other lineages were found from subtropical islands (L3) and a subarctic area (L4) of Japan. The Canadian population showed one unique haplotype (L2) diverged from the other lineages. In the combined nuclear gene tree, individuals with mitochondrial L4 haplotypes diverged from those with the other mitochondrial haplotypes L1-L3; although individuals with L1-L3 haplotypes showed shallow divergences in the nuclear gene sequences, individuals from Southeast Asia and Canada each formed a monophyletic group. Overall, the genetic composition of the Southeast Asian populations was closely related to that of temperate Japanese populations, suggesting recent gene flow between these regions. The Canadian population might have originated from anthropogenic introduction from somewhere in Asia, but the possibility that it could have spread across the Beringian land bridge cannot be ruled out.

Full Text Available The coastal mosquitoAedes togoi occurs more or less continuously from subarctic to subtropic zones along the coasts of the Japanese islands and the East Asian mainland. It occurs also in tropical Southeast Asia and the North American Pacific coast, and the populations there are thought to have been introduced from Japan by ship. To test this hypothesis, the genetic divergence among geographic populations of A. togoi was studied using one mitochondrial and three nuclear gene sequences. We detected 71 mitochondrial haplotypes forming four lineages, with high nucleotide diversity around temperate Japan and declining towards peripheral ranges. The major lineage (L1 comprised 57 haplotypes from temperate and subarctic zones in Japan and Southeast Asia including southern China and Taiwan. Two other lineages were found from subtropical islands (L3 and a subarctic area (L4 of Japan. The Canadian population showed one unique haplotype (L2 diverged from the other lineages. In the combined nuclear gene tree, individuals with mitochondrial L4 haplotypes diverged from those with the other mitochondrial haplotypes L1-L3; although individuals with L1-L3 haplotypes showed shallow divergences in the nuclear gene sequences, individuals from Southeast Asia and Canada each formed a monophyletic group. Overall, the genetic composition of the Southeast Asian populations was closely related to that of temperate Japanese populations, suggesting recent gene flow between these regions. The Canadian population might have originated from anthropogenic introduction from somewhere in Asia, but the possibility that it could have spread across the Beringian land bridge cannot be ruled out.

Full Text Available Mosquito-borne diseases such as dengue fever, chikungunya or malaria affect millions of people each year and control solutions are urgently needed. An international research program is currently being developed that relies on the introduction of the bacterial endosymbiont Wolbachia pipientis into Aedes aegypti to control dengue transmission. In order to prepare for open-field testing releases of Wolbachia-infected mosquitoes, an intensive social research and community engagement program was undertaken in Cairns, Northern Australia. The most common concern expressed by the diverse range of community members and stakeholders surveyed was the necessity of assuring the safety of the proposed approach for humans, animals and the environment. To address these concerns a series of safety experiments were undertaken. We report in this paper on the experimental data obtained, discuss the limitations of experimental risk assessment and focus on the necessity of including community concerns in scientific research.

Full Text Available Abstract Background Microsatellite markers have proven useful in genetic studies in many organisms, yet microsatellite-based studies of the dengue and yellow fever vector mosquitoAedes aegypti have been limited by the number of assayable and polymorphic loci available, despite multiple independent efforts to identify them. Here we present strategies for efficient identification and development of useful microsatellites with broad coverage across the Aedes aegypti genome, development of multiplex-ready PCR groups of microsatellite loci, and validation of their utility for population analysis with field collections from Haiti. Results From 79 putative microsatellite loci representing 31 motifs identified in 42 whole genome sequence supercontig assemblies in the Aedes aegypti genome, 33 microsatellites providing genome-wide coverage amplified as single copy sequences in four lab strains, with a range of 2-6 alleles per locus. The tri-nucleotide motifs represented the majority (51% of the polymorphic single copy loci, and none of these was located within a putative open reading frame. Seven groups of 4-5 microsatellite loci each were developed for multiplex-ready PCR. Four multiplex-ready groups were used to investigate population genetics of Aedes aegypti populations sampled in Haiti. Of the 23 loci represented in these groups, 20 were polymorphic with a range of 3-24 alleles per locus (mean = 8.75. Allelic polymorphic information content varied from 0.171 to 0.867 (mean = 0.545. Most loci met Hardy-Weinberg expectations across populations and pairwise FST comparisons identified significant genetic differentiation between some populations. No evidence for genetic isolation by distance was observed. Conclusion Despite limited success in previous reports, we demonstrate that the Aedes aegypti genome is well-populated with single copy, polymorphic microsatellite loci that can be uncovered using the strategy developed here for rapid and efficient

Background A fundamental understanding of the spatial distribution and ecology of mosquito larvae is essential for effective vector control intervention strategies. In this study, data-driven decision tree models, generalized linear models and ordination analysis were used to identify the most important biotic and abiotic factors that affect the occurrence and abundance of mosquito larvae in Southwest Ethiopia. Methods In total, 220 samples were taken at 180 sampling locations during the years 2010 and 2012. Sampling sites were characterized based on physical, chemical and biological attributes. The predictive performance of decision tree models was evaluated based on correctly classified instances (CCI), Cohen’s kappa statistic (κ) and the determination coefficient (R2). A conditional analysis was performed on the regression tree models to test the relation between key environmental and biological parameters and the abundance of mosquito larvae. Results The decision tree model developed for anopheline larvae showed a good model performance (CCI = 84 ± 2%, and κ = 0.66 ± 0.04), indicating that the genus has clear habitat requirements. Anopheline mosquito larvae showed a widespread distribution and especially occurred in small human-made aquatic habitats. Water temperature, canopy cover, emergent vegetation cover, and presence of predators and competitors were found to be the main variables determining the abundance and distribution of anopheline larvae. In contrast, anopheline mosquito larvae were found to be less prominently present in permanent larval habitats. This could be attributed to the high abundance and diversity of natural predators and competitors suppressing the mosquito population densities. Conclusions The findings of this study suggest that targeting smaller human-made aquatic habitats could result in effective larval control of anopheline mosquitoes in the study area. Controlling the occurrence of mosquito larvae via drainage

To address concerns that a flavivirus vaccine/wild-type recombinant virus might have a high mosquito infectivity phenotype, the yellow fever virus (YFV) 17D backbone of the ChimeriVax– dengue 4 virus was replaced with the corresponding gene sequences of the virulent YFV Asibi strain. Field-collected and laboratory-colonized Aedes aegypti mosquitoes were fed on blood containing each of the viruses under investigation and held for 14 days after infection. Infection and dissemination rates were ...

Full Text Available The Asian tiger mosquitoAedes (Stegomya albopictus is an invasive species that has spread across the world in the last two decades, showing a great capacity to adapt to contrasting climates and environments. While demonstrated in many insects, the contribution of bacterial symbionts in Aedes ecology is a challenging aspect that needs to be investigated however. Some bacterial species have already been identified in Ae. albopictus using classical methods, but a more accurate survey of mosquito-associated bacterial diversity is needed to decipher the potential biological functions of bacterial symbionts in mediating or constraining insect adaptation. We surveyed the bacteria associated with field populations of Ae. albopictus from Madagascar by pyrosequencing 16S rRNA gene amplicons. Different aspects of amplicon preparation and sequencing depth were tested to optimise the breadth of bacterial diversity identified. The results revealed that all mosquitoes collected from different sites have a bacterial microbiota dominated by a single taxon, Wolbachia pipientis, which accounted for about 99% of all 98,520 sequences obtained. Ae. albopictus is known to harbour two Wolbachia strains, wAlbA and wAlbB, and quantitative PCR was used to estimate the relative densities, i.e. the bacteria-to-host gene ratios, of the strains in individual mosquitoes. Relative densities were between 6.25 × 100.01 and 5.47 × 100.1 for wAlbA and between 2.03 × 100.1 and 1.4 × 101 for wAlbB. Apart from Wolbachia, a total of 32 bacterial taxa were identified at the genus level using the different in method variations. Diversity index values were low and probably underestimated the true diversity due to the high abundance of Wolbachia sequences vastly outnumbering sequences from other taxa. Further studies should implement alternative strategies to specifically discard from analysis any sequences from Wolbachia, the dominant endosymbiotic bacterium in Ae. albopictus from

Container-breeding mosquitoes, such as Aedes triseriatus, ingest biofilms and filter water column microorganisms directly to obtain the bulk of their nutrition. Scirtid beetles often co-occur with A. triseriatus and may facilitate the production of mosquito adults under low-resource conditions. Using molecular genetic techniques and quantitative assays, we observed changes in the dynamics and composition of bacterial and fungal communities present on leaf detritus and in the water column when scirtid beetles co-occur with A. triseriatus. Data from terminal restriction fragment polymorphism analysis indicated scirtid presence alters the structure of fungal communities in the water column but not leaf-associated fungal communities. Similar changes in leaf and water bacterial communities occurred in response to mosquito presence. In addition, we observed increased processing of leaf detritus, higher leaf-associated enzyme activity, higher bacterial productivity, and higher leaf-associated fungal biomass when scirtid beetles were present. Such shifts suggest beetle feeding facilitates mosquito production indirectly through the microbial community rather than directly through an increase in available fine particulate organic matter.

Insecticide resistance is a serious problem facing the effective control of insect vectors of disease. Insensitive acetylcholinesterase (AChE) confers resistance to organophosphorus (OP) and carbamate insecticides and is a widespread resistance mechanism in vector mosquitoes. Although the point mutations that underlie AChE insensitivity have been described from Drosophila, the Colorado potato beetle, and house flies, no resistance associated mutations have been documented from mosquitoes to date. We are therefore using a cloned acetylcholinesterase gene from the yellow fever mosquitoAedes aegypti as a model in which to perform site directed mutagenesis in order to understand the effects of potential resistance associated mutations. The same resistance associated amino-acid replacements as found in other insects also confer OP and carbamate resistance to the mosquito enzyme. Here we describe the levels of resistance conferred by different combinations of these mutations and the effects of these mutations on the kinetics of the AChE enzyme. Over-expression of these constructs in baculovirus will facilitate purification of each of the mutant enzymes and a more detailed analysis of their associated inhibition kinetics.

Mosquito control is facing a threat due to the emergence of resistance to synthetic insecticides. Insecticides of botanical origin may serve as suitable alternative biocontrol agents. The present study is to evaluate adulticidal activity of Clausena dentata plant extract against Aedes aegypti and Culex quinquefasciatus mosquitoes. The adult mortality was observed after 24 h of exposure. The highest mortality was found in acetone extracts against Ae. aegypti and Cx. quinquefasciatus with the LC50 and LC90 4.1783 mg/ml (3.8201-7.1026), 9.3884 mg/ml (7. 8258-13.1820) and 4.2451 mg/ml (3.8547-8.0254), 12.3214 mg/ml (10.9287-16.2220), respectively. Smoke toxicity was observed at 10-min interval for 40 min, and the mortality data were recorded. Result shows that Ae. aegypti and Cx. quinquefasciatus are 85 ± 2 and 89 ± 1.5, respectively. A mortality of 100 % was recorded in the commercial mosquito control. These results suggest that the leaf extracts of C. dentata have a potential to be used as an ideal eco-friendly approach for the control of mosquitoes.

Susceptibility of the mosquitoAedes aegypti to the malarial parasite Plasmodium gallinaceum was investigated as a quantitative trait using restriction fragment length polymorphisms (RFLP). Two F{sub 2} populations of mosquitoes were independently prepared from pairwise matings between a highly susceptible and a refractory strain of A. aegypti. RFLP were tested for association with oocyst development on the mosquito midgut. Two putative quantitative trait loci (QTL) were identified that significantly affect susceptibility. One QTL, pgs [2,LF98], is located on chromosome 2 and accounted for 65 and 49% of the observed phenotypic variance in the two populations, respectively. A second QTL, pgs[3,MalI], is located on chromosome 3 and accounted for 14 and 10% of the observed phenotypic variance in the two populations, respectively. Both QTL exhibit a partial dominance effect on susceptibility, wherein the dominance effect is derived from the refractory parent. No indication of epistasis between these QTL was detected. Evidence suggests that either a tightly linked cluster of independent genes or a single locus affecting susceptibility to various mosquito-borne parasites and pathogens has evolved near the LF98 locus; in addition to P. gallinaceum susceptibility, this general genome region has previously been implicated in susceptibility to the filaria nematode Brugia malayi and the yellow fever virus. 35 refs., 2 figs., 3 tabs.

Full Text Available BACKGROUND: The tiger mosquito (Aedes albopictus, vector of several emerging diseases, is expanding into more northerly latitudes as well as into higher altitudes in northern Italy. Changes in the pattern of distribution of the tiger mosquito may affect the potential spread of infectious diseases transmitted by this species in Europe. Therefore, predicting suitable areas of future establishment and spread is essential for planning early prevention and control strategies. METHODOLOGY/PRINCIPAL FINDINGS: To identify the areas currently most suitable for the occurrence of the tiger mosquito in the Province of Trento, we combined field entomological observations with analyses of satellite temperature data (MODIS Land Surface Temperature: LST and human population data. We determine threshold conditions for the survival of overwintering eggs and for adult survival using both January mean temperatures and annual mean temperatures. We show that the 0°C LST threshold for January mean temperatures and the 11°C threshold for annual mean temperatures provide the best predictors for identifying the areas that could potentially support populations of this mosquito. In fact, human population density and distance to human settlements appear to be less important variables affecting mosquito distribution in this area. Finally, we evaluated the future establishment and spread of this species in relation to predicted climate warming by considering the A2 scenario for 2050 statistically downscaled at regional level in which winter and annual temperatures increase by 1.5 and 1°C, respectively. CONCLUSIONS/SIGNIFICANCE: MODIS satellite LST data are useful for accurately predicting potential areas of tiger mosquito distribution and for revealing the range limits of this species in mountainous areas, predictions which could be extended to an European scale. We show that the observed trend of increasing temperatures due to climate change could facilitate further

Obligatory blood-triggered reproductive strategy is an evolutionary adaptation of mosquitoes for rapid egg development. It contributes to the vectorial capacity of these insects. Therefore, understanding the molecular mechanisms underlying reproductive processes is of particular importance. Here, we report that microRNA-309 (miR-309) plays a critical role in mosquito reproduction. A spatiotemporal expression profile of miR-309 displayed its blood feeding-dependent onset and ovary-specific manifestation in female Aedes aegypti mosquitoes. Antagomir silencing of miR-309 impaired ovarian development and resulted in nonsynchronized follicle growth. Furthermore, the genetic disruption of miR-309 by CRISPR/Cas9 system led to the developmental failure of primary follicle formation. Examination of genomic responses to miR-309 depletion revealed that several pathways associated with ovarian development are down-regulated. Comparative analysis of genes obtained from the high-throughput RNA sequencing of ovarian tissue from the miR-309 antagomir-silenced mosquitoes with those from the in silico computation target prediction identified that the gene-encoding SIX homeobox 4 protein (SIX4) is a putative target of miR-309. Reporter assay and RNA immunoprecipitation confirmed that SIX4 is a direct target of miR-309. RNA interference of SIX4 was able to rescue phenotypic manifestations caused by miR-309 depletion. Thus, miR-309 plays a critical role in mosquito reproduction by targeting SIX4 in the ovary and serves as a regulatory switch permitting a stage-specific degradation of the ovarian SIX4 mRNA. In turn, this microRNA (miRNA)-targeted degradation is required for appropriate initiation of a blood feeding-triggered phase of ovarian development, highlighting involvement of this miRNA in mosquito reproduction. PMID:27489347

Full Text Available Background: Dengue Hemorrhagic Fever (DHF is a contagious disease caused by the dengue virus infection through Aedes aegypti mosquito. DHF often causes epidemics and Extraordinary Events (EE in tropical and subtropical regions, including Indonesia. Objective: This study aims to examine the relationship between knowledge and society action on the prevention of dengue with the amount of mosquitoes trapped in ovitrap in Kandai, Kendari, Indonesia. Methods: The research method was an observational study with cross sectional design. The research was conducted at Kandai village, Kendari on May to July 2014. It was 87 homes with 87 families were recruited as sample size for this study. Ovitrap was used inside and outside of their homes. Results: DCB family knowledge and the existence of Aedes sp mosquitoes eggs on ovitrap were analyzed and observed in this study. Chi square test showed that X2 count value (1.261 was less than X2 table value (3.814 at error level 5% (α = 0.05 indicated that Ha was rejected and Ho was accepted. Meanwhile, Chi square test for the action of the family about DCB and the existence of eggs Aedes sp mosquito on ovitrap showed that X2 count value (4.115 was greater than X2 table value (3.814 at the error level 5% (α = 0.05, indicated that Ho was rejected and Ha was accepted. Conclusions: It was concluded that (1 there was no relationship between DCB family knowledge and the existence of Aedes sp mosquitoes eggs on ovitrap, and (2 there was a relationship between the action of family about DCB and the existence of aedes sp mosquitoes eggs on ovitrap in Kandai Kendari.

Full Text Available BACKGROUND: New strategies to eliminate dengue have been proposed that specifically target older Aedes aegypti mosquitoes, the proportion of the vector population that is potentially capable of transmitting dengue viruses. Evaluation of these strategies will require accurate and high-throughput methods of predicting mosquito age. We previously developed an age prediction assay for individual Ae. aegypti females based on the transcriptional profiles of a selection of age responsive genes. Here we conducted field testing of the method on Ae. aegypti that were entirely uncaged and free to engage in natural behavior. METHODOLOGY/PRINCIPAL FINDINGS: We produced "free-range" test specimens by releasing 8007 adult Ae. aegypti inside and around an isolated homestead in north Queensland, Australia, and recapturing females at two day intervals. We applied a TaqMan probe-based assay design that enabled high-throughput quantitative RT-PCR of four transcripts from three age-responsive genes and a reference gene. An age prediction model was calibrated on mosquitoes maintained in small sentinel cages, in which 68.8% of the variance in gene transcription measures was explained by age. The model was then used to predict the ages of the free-range females. The relationship between the predicted and actual ages achieved an R(2 value of 0.62 for predictions of females up to 29 days old. Transcriptional profiles and age predictions were not affected by physiological variation associated with the blood feeding/egg development cycle and we show that the age grading method could be applied to differentiate between two populations of mosquitoes having a two-fold difference in mean life expectancy. CONCLUSIONS/SIGNIFICANCE: The transcriptional profiles of age responsive genes facilitated age estimates of near-wild Ae. aegypti females. Our age prediction assay for Ae. aegypti provides a useful tool for the evaluation of mosquito control interventions against dengue where

Full Text Available Immatures of both Aedes aegypti and Aedes albopictus have been found in water-holding bromeliad axils in Brazil. Removal of these plants or their treatment with insecticides in public and private gardens have been undertaken during dengue outbreaks in Brazil despite uncertainty as to their importance as productive habitats for dengue vectors. From March 2005-February 2006, we sampled 120 randomly selected bromeliads belonging to 10 species in a public garden less than 200 m from houses in a dengue-endemic neighborhood in Rio de Janeiro. A total of 2,816 mosquito larvae and pupae was collected, with an average of 5.87 immatures per plant per collection. Culex (Microculex pleuristriatus and Culex spp of the Ocellatus Group were the most abundant culicid species, found in all species of bromeliads; next in relative abundance were species of the genus Wyeomyia. Only two individuals of Ae. aegypti (0.07% and five of Ae. albopictus(0.18% were collected from bromeliads. By contrast, immatures of Ae. aegypti were found in manmade containers in nearly 5% of nearby houses. These results demonstrate that bromeliads are not important producers of Ae. aegypti and Ae. albopictus and, hence, should not be a focus for dengue control. However, the results of this study of only one year in a single area may not represent outcomes in other urban localities where bromeliads, Ae. aegypti and dengue coincide in more disturbed habitats.

We conducted this study to assess how well a pupal survey of dengue mosquito vectors, Aedes aegypti and Aedes albopictus, is able to target the most productive breeding sites. The study was carried out monthly during the rainy season (8 months) in 2008 in Cuba, Cebu City, Philippines. The hypotheses tested were: 1) most pupae of Ae. aegypti or Ae. albopictus were produced in a few types of breeding sites and 2) the most productive types of breeding sites for each species were the most abundant. Approximately 2,500 pupae were collected from 554 breeding sites in 279 houses. Thirty-eight point four percent of ten types of breeding sites were positive for Ae. aegypti, and 11.9% of nine types of sites were positive for Ae. albopictus. Plastic drums (40.2%), metal drums (29.6%), and plastic containers (10.5%) were the key sites for Ae. aegypti pupae, whereas bamboo stumps (28.5%), plastic drums (21.1%), and rubber tires (19.1%) were the key sites for Ae. albopictus. The most productive breeding sites for Ae. aegypti were common but not the most common for Ae. albopictus. These results are relevant for dengue vector control programs.

Full Text Available Aedes albopictus is a major vector of dengue and Chikungunya viruses. Olfaction plays a vital role in guiding mosquito behaviors and contributes to their ability to transmit pathogens. Odorant-binding proteins (OBPs are abundant in insect olfactory tissues and involved in the first step of odorant reception. While comprehensive descriptions are available of OBPs from Aedes aegypti, Culex quinquefasciatus and Anopheles gambiae, only a few genes from Ae. albopictus have been reported. In this study, twenty-one putative AalbOBP genes were cloned using their homologues in Ae. aegypti to query an Ae. albopictus partial genome sequence. Two antenna-specific OBPs, AalbOBP37 and AalbOBP39, display a remarkable similarity in their overall folding and binding pockets, according to molecular modeling. Binding affinity assays indicated that AalbOBP37 and AalbOBP39 had overlapping ligand affinities and are affected in different pH condition. Electroantennagrams (EAG and behavioral tests show that these two genes were involved in olfactory reception. An improved understanding of the Ae. albopictus OBPs is expected to contribute to the development of more efficient and environmentally-friendly mosquito control strategies.

Full Text Available Background: Considering the disadvantages of chemical insecticides, we aimed to evaluate Vorticella parasites for control of mosquito larvae of Anopheles stephensi and Aedes aegypti at different larval stages.Methods: Vorticella sp infected mosquito larvae were crushed in the 0.85% saline and homogenized well to get Vorti­cella in suspension. The effects of Vorticella sp infections on larval development were investigated by inoculat­ing protozoan on different larval instars of An. stephensi and Ae. aegypti and observed under light microscope. Le­thal time of the Vorticella infected larvae at different stages was calculated.Results: First and 2nd larval instars of both An. stephensi and Ae. aegypti did not show signs of infection by Vorti­cella sp., whereas 3rd instars of An. stephensi showed more Vorticella infection than those of Ae. aegypti. However, 4th larval instars of both mosquitoes were heavily infected with Vorticella parasite which was responsible for slug­gish movements of larvae and eventually death. Moreover, parasites (Vorticella spp were responsible for more than 90% reduction in adult emergence for both infected An. stephensi and Ae. aegypti.Conclusion: This study provides insights for mosquito larvicidal action of surface parasite Vorticella on different larval stages of An. stephensi and Ae. Aegypti. It could be suggested as a potential candidate in mosquito biocontrol programs.

Wolbachia is a bacterial endosymbiont that naturally infects a wide range of insect species, and causes drastic changes to host biology. Stable infections of Wolbachia in mosquitoes can inhibit infection with medically important pathogens such as dengue virus and malaria-causing Plasmodium parasites. However, some native Wolbachia strains can enhance infection with certain pathogens, as is the case for the mosquitoAedes fluviatilis, where infection with Plasmodium gallinaceum is enhanced by the native wFlu Wolbachia strain. To better understand the biological interactions between mosquitoes and native Wolbachia infections, and to investigate the process of pathogen enhancement, we used RNA-Seq to generate the transcriptome of Ae. fluviatilis with and without Wolbachia infection. In total, we generated 22,280,160 Illumina paired-end reads from Wolbachia-infected and uninfected mosquitoes, and used these to make a de novo transcriptome assembly, resulting in 58,013 contigs with a median sequence length of 443 bp and an N50 of 2454 bp. Contigs were annotated through local alignments using BlastX, and associated with both gene ontology and KEGG orthology terms. Through baySeq, we identified 159 contigs that were significantly upregulated due to Wolbachia infection, and 98 that were downregulated. Critically, we saw no changes to Toll or IMD immune gene transcription, but did see evidence that wFlu infection altered the expression of several bacterial recognition genes, and immune-related genes that could influence Plasmodium infection. wFlu infection also had a widespread effect on a number of host physiological processes including protein, carbohydrate and lipid metabolism, and oxidative stress. We then compared our data set with transcriptomic data for other Wolbachia infections in Aedes aegypti, and identified a core set of 15 gene groups associated with Wolbachia infection in mosquitoes. While the scale of transcriptional changes associated with wFlu infection

Aedes aegypti (L.) (Diptera: Culicidae), the main vector of dengue and urban yellow fever in the world, is highly adapted to the human environment. Artificial containers are the most common larval habitat for the species, but it may develop in tree holes and other phytotelmata. This study assessed whether tree holes in San Ramón de la Nueva Orán, a city located in subtropical montane moist forest where dengue outbreaks occur, are relevant as larval habitat for Ae. aegypti and if the species may be found in natural areas far from human habitations. Water holding tree holes were sampled during 3 years once a month along the rainy season using a siphon bottle, in urban and suburban sites within the city and in adjacent forested areas. Larvae and pupae were collected and the presence and volume of water in each tree hole were recorded. Finding Ae. aegypti in forested areas was an isolated event; however, the species was frequently collected from tree holes throughout the city and along the sampling period. Moreover, larvae were collected in considerably high numbers, stressing the importance of taking into account these natural cavities as potential reinfestation foci within dengue control framework.

Background An in depth understanding of mosquito breeding biology and factors regulating population sizes is fundamental for vector population control. This paper presents results from a survey of mosquito breeding habitats and the efficacy of neem chippings as a potential larvicide that can be integrated in mosquito control on Nyabondo Plateau in western Kenya. Results Six main mosquito habitat types namely artificial ponds, abandoned fish ponds, active fish ponds, open drains, temporary poo...

Full Text Available BACKGROUND: The Asian tiger mosquito, Aedes albopictus (Skuse, is a vector of several arboviruses including dengue and chikungunya. This highly invasive species originating from Southeast Asia has travelled the world in the last 30 years and is now established in Europe, North and South America, Africa, the Middle East and the Caribbean. In the absence of vaccine or antiviral drugs, efficient mosquito control strategies are crucial. Conventional control methods have so far failed to control Ae. albopictus adequately. METHODOLOGY/PRINCIPAL FINDINGS: Germline transformation of Aedes albopictus was achieved by micro-injection of embryos with a piggyBac-based transgene carrying a 3xP3-ECFP marker and an attP site, combined with piggyBac transposase mRNA and piggyBac helper plasmid. Five independent transgenic lines were established, corresponding to an estimated transformation efficiency of 2-3%. Three lines were re-injected with a second-phase plasmid carrying an attB site and a 3xP3-DsRed2 marker, combined with PhiC31 integrase mRNA. Successful site-specific integration was observed in all three lines with an estimated transformation efficiency of 2-6%. CONCLUSIONS/SIGNIFICANCE: Both piggybac- and site-specific PhiC31-mediated germline transformation of Aedes albopictus were successfully achieved. This is the first report of Ae. albopictus germline transformation and engineering, a key step towards studying and controlling this species using novel molecular techniques and genetic control strategies.

An entomological survey of Aedesmosquitoes was initiated at the container terminal of the Autonomous Port of Abidjan in 2009 following the occurrence of two outbreaks of yellow fever in less than 10 years and dengue cases reported in 2008 among expatriates returning from Côte d’Ivoire (Ivory Coast). It was based on regular monitoring of ovitraps from July 2009 to June 2010. A total of 4,739 adult mosquitoes represented by four species of Aedes (97% of total) and one unexpected species of Culex (3%) were obtained. Aedes aegypti was dominant with 98% of total Aedes (n = 4,594). Its density variation was closely related to the amount of rainfall. The other species of Aedes were collected in the second half of the major rainy season including Ae. albopictus (1.17% of Aedes) and Ae. angustus (0.13%) whose presence was discovered for the first time in Côte d’Ivoire. PMID:23567057

Full Text Available Behaviour comparisons of Aedes scapularis and Ae. serratus are presented. Results were obtained by sampling Aedes adult mosquitoes at several places in the rural anthropic environment in the Ribeira Valley region of S. Paulo State, Brazil. Aedes dominance was shared by those two species, but Ae. scapularis Sshowed a clear tendency to frequent the modified environment, while Ae. serratus was to be found in the more preserved ones, here represented by the vestigial patchy forests. Regarding the open cultivated land and the dwelling environments, Ae. scapularis preponderates. Considering the regional developmental phases, this mosquito showed a remarkable increase in the modified environment differently from Ae. serratus that underwent a considerable decrease in migrating from the forest to the anthropic environment. As a consequence of these results it is reasonable to conclude that Ae. scapularis may be considered as an epidemiologically efficient vector and that it quite probably played this role in the Rocio encephalitis and other arbovirus epidemics.

Full Text Available Behaviour comparisons of Aedes scapularis and Ae. serratus are presented. Results were obtained by sampling Aedes adult mosquitoes at several places in the rural anthropic environment in the Ribeira Valley region of S. Paulo State, Brazil. Aedes dominance was shared by those two species, but Ae. scapularis Sshowed a clear tendency to frequent the modified environment, while Ae. serratus was to be found in the more preserved ones, here represented by the vestigial patchy forests. Regarding the open cultivated land and the dwelling environments, Ae. scapularis preponderates. Considering the regional developmental phases, this mosquito showed a remarkable increase in the modified environment differently from Ae. serratus that underwent a considerable decrease in migrating from the forest to the anthropic environment. As a consequence of these results it is reasonable to conclude that Ae. scapularis may be considered as an epidemiologically efficient vector and that it quite probably played this role in the Rocio encephalitis and other arbovirus epidemics.

The genetic manipulation of mosquito vectors is an alternative strategy in the fight against malaria. It was previously shown that bee venom phospholipase A2 (PLA2) inhibits ookinete invasion of the mosquito midgut although mosquito fitness was reduced. To maintain the PLA2 blocking ability without compromising mosquito biology, we mutated the protein-coding sequence to inactivate the enzyme while maintaining the protein’s structure. DNA encoding the mutated PLA2 (mPLA2) was placed downstream...

In Cairns, Australia, the impacts on Aedes aegypti L. (Diptera: Culicidae) populations of two types of 'lure & kill' (L&K) lethal ovitraps (LOs), the standard lethal ovitrap (SLO) and the biodegradable lethal ovitrap (BLO) were measured during three mass-trapping interventions. To assess the efficacy of the SLO, two interventions (one dry season and one wet season) were conducted in three discrete areas, each lasting 4 weeks, with the following treatments: (i) SLOs (>200 traps, approximately 4/premise), BG-sentinel traps (BGSs; approximately 15, 1/premise) and larval control (container reduction and methoprene treatment) and (ii) larval control alone, and (iii) untreated control. Female Ae. aegypti populations were monitored for 4 weeks pre- and post-treatment in all three areas using BGSs and sticky ovitraps (SOs) or non-lethal regular ovitraps (ROs). In the dry season, 206 SLOs and 15 BGSs set at 54 and 15 houses, respectively, caught and killed an estimated 419 and 73 female Ae. aegypti, respectively. No significant decrease in collection size of female Ae. aegypti could be attributed to the treatments. In the wet season, 243 SLOs and 15 BGSs killed approximately 993 and 119 female Ae. aegypti, respectively. The mean number of female Ae. aegypti collected after 4 weeks with SOs and BGSs was significantly less than the control (LSD post-hoc test). The third mass-trapping intervention was conducted using the BLO during the wet season in Cairns. For this trial, three treatment areas were each provided with BLOs (>500, approximately 4/premise) plus larval control, and an untreated control area was designated. Adult female Ae. aegypti were collected for 4 weeks pre- and post-treatment using 15 BGSs and 20 SOs. During this period, 53.2% of BLOs contained a total of 6654 Ae. aegypti eggs. Over the intervention period, collections of Ae. aegypti in the treatment areas were significantly less than in the control area for BGSs but not SOs. An influx of relatively large

Conclusions: Ae. albopictus is a well-known candidate for dengue transmission in Japan. However, our data suggest that Ae. f. miyarai from Ishigaki Island (near Okinawa Island and Ae. galloisi from Hokkaido (Northern Japan should also be regarded as potential vectors for dengue transmission in these regions. Further studies on these mosquitoes should be conducted.

Dengue viruses (DENV) are comprised in four related serotypes (DENV-1 to 4) and are critically important arboviral pathogens affecting human populations in the tropics. South American countries have seen the reemergence of DENV since the 1970's associated with the progressive re-infestation by the mosquito vector, Aedes aegypti. In French Guiana, DENV is now endemic with the co-circulation of different serotypes resulting in viral epidemics. Between 2009 and 2010, a predominant serotype change occurred from DENV-1 to DENV-4 suggesting a competitive displacement. The aim of the present study was to evaluate the potential role of the mosquito in the selection of the new epidemic serotype. To test this hypothesis of competitive displacement of one serotype by another in the mosquito vector, we performed mono- and co-infections of local Ae. aegypti collected during the inter-epidemic period with both viral autochthonous epidemic serotypes and compared infection, dissemination and transmission rates. We performed oral artificial infections of F1 populations in BSL-3 conditions and analyzed infection, dissemination and transmission rates. When two populations of Ae. aegypti from French Guiana were infected with either serotype, no significant differences in dissemination and transmission were observed between DENV-1 and DENV-4. However, in co-infection experiments, a strong competitive advantage for DENV-4 was seen at the midgut level leading to a much higher dissemination of this serotype. Furthermore only DENV-4 was present in Ae. aegypti saliva and therefore able to be transmitted. In an endemic context, mosquito vectors may be infected by several DENV serotypes. Our results suggest a possible competition between serotypes at the midgut level in co-infected mosquitoes leading to a drastically different transmission potential and, in this case, favoring the competitive displacement of DENV-1 by DENV-4. This phenomenon was observed despite a similar replicative fitness

Effective mosquito control is vital to curtail the devastating health effects of many vectored diseases. RNA interference (RNAi)-mediated control of mosquitoes is an attractive alternative to conventional chemical pesticides. Previous studies have suggested that transcripts for inhibitors of apoptosis (IAPs) may be good RNAi targets. To revisit and extend previous reports, we examined the expression of Aedes aegypti (L.) IAPs (AaeIAPs) 1, 2, 5, 6, 9, and a viral IAP-associated factor (vIAF) as well as Anopheles quadrimaculatus Say and Culex quinquefasciatus Say IAP1 homologs (AquIAP1 and CquIAP1) in adult females. Expression profiles of IAPs suggested that some older female mosquitoes had significantly higher IAP mRNA levels when compared to the youngest ones. Minor differences in expression of AaeIAPs were observed in mosquitoes that imbibed a bloodmeal, but the majority of the time points (up to 48 h) were not significantly different. Although in vitro experiments with the Ae. aegypti Aag-2 cell line demonstrated that the various AaeIAPs could be effectively knocked down within one day after dsRNA treatment, only Aag-2 cells treated with dsIAP1 displayed apoptotic morphology. Gene silencing and mortality were also evaluated after topical application and microinjection of the same dsRNAs into female Ae. aegypti. In contrast to previous reports, topical administration of dsRNA against AaeIAP1 did not yield a significant reduction in gene expression or increased mortality. Knockdown of IAP1 and other IAPs by microinjection did not result in significant mortality. In toto, our findings suggest that IAPs may not be suitable RNAi targets for controlling adult mosquito populations.

Only adult female mosquitoes feed on blood, while both genders take sugar meals. Accordingly, several compounds associated with blood feeding (i.e. vasodilators, anti-clotting, anti-platelets) are found only in female glands, while enzymes associated with sugar feeding or antimicrobials (such as lysozyme) are found in the glands of both sexes. We performed de novo assembly of reads from adult Aedes aegypti female and male salivary gland libraries (285 and 90 million reads, respectively). By mapping back the reads to the assembled contigs, plus mapping the reads from a publicly available Ae. aegypti library from adult whole bodies, we identified 360 transcripts (including splice variants and alleles) overexpressed tenfold or more in the glands when compared to whole bodies. Moreover, among these, 207 were overexpressed fivefold or more in female vs. male salivary glands, 85 were near equally expressed and 68 were overexpressed in male glands. We call in particular the attention to C-type lectins, angiopoietins, female-specific Antigen 5, the 9.7 kDa, 12-14 kDa, 23.5 kDa, 62/34 kDa, 4.2 kDa, proline-rich peptide, SG8, 8.7 kDa family and SGS fragments: these polypeptides are all of unknown function, but due to their overexpression in female salivary glands and putative secretory nature they are expected to affect host physiology. We have also found many transposons (some of which novel) and several endogenous viral transcripts (probably acquired by horizontal transfer) which are overexpressed in the salivary glands and may play some role in tissue-specific gene regulation or represent a mechanism of virus interference. This work contributes to a near definitive catalog of male and female salivary gland transcripts from Ae. aegypti, which will help to direct further studies aiming at the functional characterization of the many transcripts with unknown function and the understanding of their role in vector-host interaction and pathogen transmission.

Full Text Available Only adult female mosquitoes feed on blood, while both genders take sugar meals. Accordingly, several compounds associated with blood feeding (i.e. vasodilators, anti-clotting, anti-platelets are found only in female glands, while enzymes associated with sugar feeding or antimicrobials (such as lysozyme are found in the glands of both sexes. We performed de novo assembly of reads from adult Aedes aegypti female and male salivary gland libraries (285 and 90 million reads, respectively. By mapping back the reads to the assembled contigs, plus mapping the reads from a publicly available Ae. aegypti library from adult whole bodies, we identified 360 transcripts (including splice variants and alleles overexpressed tenfold or more in the glands when compared to whole bodies. Moreover, among these, 207 were overexpressed fivefold or more in female vs. male salivary glands, 85 were near equally expressed and 68 were overexpressed in male glands. We call in particular the attention to C-type lectins, angiopoietins, female-specific Antigen 5, the 9.7 kDa, 12-14 kDa, 23.5 kDa, 62/34 kDa, 4.2 kDa, proline-rich peptide, SG8, 8.7 kDa family and SGS fragments: these polypeptides are all of unknown function, but due to their overexpression in female salivary glands and putative secretory nature they are expected to affect host physiology. We have also found many transposons (some of which novel and several endogenous viral transcripts (probably acquired by horizontal transfer which are overexpressed in the salivary glands and may play some role in tissue-specific gene regulation or represent a mechanism of virus interference. This work contributes to a near definitive catalog of male and female salivary gland transcripts from Ae. aegypti, which will help to direct further studies aiming at the functional characterization of the many transcripts with unknown function and the understanding of their role in vector-host interaction and pathogen transmission.

Aedes species mosquitoes transmit chikungunya virus, as well as dengue and Zika viruses, and bite most often during the day.* Infectious mosquito bites frequently occur in and around homes (1,2). Caribbean countries first reported local transmission of chikungunya virus in December 2013, and soon after, chikungunya virus spread throughout the Americas (3). Puerto Rico reported its first laboratory-positive chikungunya case in May 2014 (4), and subsequently identified approximately 29,000 suspected cases throughout the island by the end of 2015.(†) Because conventional vector control approaches often fail to result in effective and sustainable prevention of infection with viruses transmitted by Aedesmosquitoes (5), and to improve surveillance of mosquito population densities, CDC developed an Autocidal Gravid Ovitrap (AGO) (6) to attract and capture the female Aedes aegypti mosquitoes responsible for transmission of infectious agents to humans (Figure). The AGO trap is a simple, low-cost device that requires no use of pesticides and no servicing for an extended period of time (6).

We present a mathematical model to describe the dynamics of mosquito population when sterile male mosquitoes (produced by irradiation) are introduced as a biological control, besides the application of insecticide. In order to analyze the minimal effort to reduce the fertile female mosquitoes, we search for the optimal control considering the cost of insecticide application, the cost of the production of irradiated mosquitoes and their delivery as well as the social cost (proportional to the number of fertilized females mosquitoes). The optimal control is obtained by applying the Pontryagin's Maximum Principle. Copyright 2009 Elsevier Inc. All rights reserved.

Full Text Available Abstract Background Resistance to chemical insecticides plus high morbidity rates have lead to rising interest in fungi as candidates for biocontrol agents of mosquito vectors. In most studies fungal infections have been induced by exposure of mosquitoes to various surfaces treated with conidia. In the present study eight Mexican strains of Beauveria bassiana were assessed against Aedes aegypti by direct exposure of females to 6 × 108 conidia ml -1 on a filter paper, afterwards, the transmission of the least and most virulent isolates was evaluated by mating behavior from virgin, fungus-contaminated male to females, to examine this ethological pattern as a new approach to deliver conidia against the dengue vector. Methods In an exposure chamber with a filter paper impregnated with 6 × 108 conidia ml -1 of the least and most virulent strains of B. bassiana, 6-8 day old males of A. aegypti were exposed for 48 hours, and then transferred individually (each one was a replicate to another chamber and confined with twenty healthy females of the same age. Clean males were used in controls. Survival, infection by true mating (insemination or by mating attempts (no insemination and fecundity were daily registered until the death of last female. Data analysis was conducted with proc glm for unbalanced experiments and means were separated with the Ryan test with SAS. Results All strains were highly virulent with LT50 ranging from 2.70 (± 0.29 to 5.33 (± 0.53 days. However the most (Bb-CBG2 and least virulent (Bb-CBG4 isolates were also transmitted by mating behavior; both killed 78-90% of females in 15 days after being confined with males that had previously been exposed for 48 hours to fungi. Of these mortality rates, 23 and 38% respectively, were infections acquired by copulations where insemination occurred. The LT50 for sexually-infected females were 7.92 (± 0.46 and 8.82 (± 0.45 days for both strains, while the one in control was 13.92 (± 0

Resistance to chemical insecticides plus high morbidity rates have lead to rising interest in fungi as candidates for biocontrol agents of mosquito vectors. In most studies fungal infections have been induced by exposure of mosquitoes to various surfaces treated with conidia. In the present study eight Mexican strains of Beauveria bassiana were assessed against Aedes aegypti by direct exposure of females to 6 × 10(8) conidia ml (-1) on a filter paper, afterwards, the transmission of the least and most virulent isolates was evaluated by mating behavior from virgin, fungus-contaminated male to females, to examine this ethological pattern as a new approach to deliver conidia against the dengue vector. In an exposure chamber with a filter paper impregnated with 6 × 10(8) conidia ml (-1) of the least and most virulent strains of B. bassiana, 6-8 day old males of A. aegypti were exposed for 48 hours, and then transferred individually (each one was a replicate) to another chamber and confined with twenty healthy females of the same age. Clean males were used in controls. Survival, infection by true mating (insemination) or by mating attempts (no insemination) and fecundity were daily registered until the death of last female. Data analysis was conducted with proc glm for unbalanced experiments and means were separated with the Ryan test with SAS. All strains were highly virulent with LT(50) ranging from 2.70 (± 0.29) to 5.33 (± 0.53) days. However the most (Bb-CBG2) and least virulent (Bb-CBG4) isolates were also transmitted by mating behavior; both killed 78-90% of females in 15 days after being confined with males that had previously been exposed for 48 hours to fungi. Of these mortality rates, 23 and 38% respectively, were infections acquired by copulations where insemination occurred. The LT(50) for sexually-infected females were 7.92 (± 0.46) and 8.82 (± 0.45) days for both strains, while the one in control was 13.92 (± 0.58). Likewise, fecundity decreased by

Full Text Available ABSTRACTINTRODUCTION:The mosquitoAedes aegypti has evolved resistance to pyrethroid insecticides. The present study evaluated Ae. aegypti from Goiânia for the resistant phenotype and for mutations associated with resistance.METHODS:Insecticide dose-response bioassays were conducted on mosquitoes descended from field-collected eggs, and polymerase chain reaction (PCR was used to genotype 90 individuals at sites implicated in pyrethroid resistance.RESULTS:All mosquito populations displayed high levels of resistance to deltamethrin, as well as high frequencies of the 1016Ile kdr and 1534Cys kdrmutations.CONCLUSIONS:Aedes aegypti populations in the Goiânia area are highly resistant to deltamethrin, presumably due to high frequencies of kdr(knockdown-resistance mutations.

The Asian tiger mosquito, Aedes albopictus, is a highly successful invasive species that transmits a number of human viral diseases, including dengue and Chikungunya fevers. This species has a large genome with significant population-based size variation. The complete genome sequence was determined for the Foshan strain, an established laboratory colony derived from wild mosquitoes from southeastern China, a region within the historical range of the origin of the species. The genome comprises 1,967 Mb, the largest mosquito genome sequenced to date, and its size results principally from an abundance of repetitive DNA classes. In addition, expansions of the numbers of members in gene families involved in insecticide-resistance mechanisms, diapause, sex determination, immunity, and olfaction also contribute to the larger size. Portions of integrated flavivirus-like genomes support a shared evolutionary history of association of these viruses with their vector. The large genome repertory may contribute to the adaptability and success of Ae. albopictus as an invasive species.

We previously reported that mosquito densoviruses (MDVs) are potential vectors for delivering foreign nucleic acids into mosquito cells. However, considering existing expression strategies, recombinant viruses would inevitably become replication-defective viruses and lose their ability for secondary transmission. The packaging limitations of the virion represent a barrier for the development of MDVs for viral paratransgenesis or as high-efficiency bioinsecticides. Herein, we report the development of a non-defective recombinant Aedes aegypti densovirus (AaeDV) miRNA expression system, mediated by an artificial intron, using an intronic miRNA expression strategy. We demonstrated that this recombinant vector could be used to overexpress endogenous miRNAs or to decrease endogenous miRNAs by generating antisense sponges to explore the biological functions of miRNAs. In addition, the vector could express antisense-miRNAs to induce efficient gene silencing in vivo and in vitro. The recombinant virus effectively self-replicated and retained its secondary transmission ability, similar to the wild-type virus. The recombinant virus was also genetically stable. This study demonstrated the first construction of a non-defective recombinant MDV miRNA expression system, which represents a tool for the functional analysis of mosquito genes and lays the foundation for the application of viral paratransgenesis for dengue virus control. PMID:26879823

The 14-3-3 proteins are evolutionarily conserved acidic proteins that form a family with several isoforms in many cell types of plants and animals. In invertebrates, including dipteran and lepidopteran insects, only two isoforms have been reported. 14-3-3 proteins are scaffold molecules that form homo- or heterodimeric complexes, acting as molecular adaptors mediating phosphorylation-dependent interactions with signaling molecules involved in immunity, cell differentiation, cell cycle, proliferation, apoptosis, and cancer. Here, we describe the presence of two isoforms of 14-3-3 in the mosquitoAedes aegypti, the main vector of dengue, yellow fever, chikungunya, and zika viruses. Both isoforms have the conserved characteristics of the family: two protein signatures (PS1 and PS2), an annexin domain, three serine residues, targets for phosphorylation (positions 58, 184, and 233), necessary for their function, and nine alpha helix-forming segments. By sequence alignment and phylogenetic analysis, we found that the molecules correspond to Ɛ and ζ isoforms (Aeae14-3-3ε and Aeae14-3-3ζ). The messengers and protein products were present in all stages of the mosquito life cycle and all the tissues analyzed, with a small predominance of Aeae14-3-3ζ except in the midgut and ovaries of adult females. The 14-3-3 proteins in female midgut epithelial cells were located in the cytoplasm. Our results may provide insights to further investigate the functions of these proteins in mosquitoes.

We previously reported that mosquito densoviruses (MDVs) are potential vectors for delivering foreign nucleic acids into mosquito cells. However, considering existing expression strategies, recombinant viruses would inevitably become replication-defective viruses and lose their ability for secondary transmission. The packaging limitations of the virion represent a barrier for the development of MDVs for viral paratransgenesis or as high-efficiency bioinsecticides. Herein, we report the development of a non-defective recombinant Aedes aegypti densovirus (AaeDV) miRNA expression system, mediated by an artificial intron, using an intronic miRNA expression strategy. We demonstrated that this recombinant vector could be used to overexpress endogenous miRNAs or to decrease endogenous miRNAs by generating antisense sponges to explore the biological functions of miRNAs. In addition, the vector could express antisense-miRNAs to induce efficient gene silencing in vivo and in vitro. The recombinant virus effectively self-replicated and retained its secondary transmission ability, similar to the wild-type virus. The recombinant virus was also genetically stable. This study demonstrated the first construction of a non-defective recombinant MDV miRNA expression system, which represents a tool for the functional analysis of mosquito genes and lays the foundation for the application of viral paratransgenesis for dengue virus control.

indoor residual applications, insecticide-treated materials (ITMs) such as cur- tains and bed nets, and the application of larvicides to aquatic...and posttreatment periods we ran 3 trials in each avenue. After each test, all deployment, collecting, and monitoring materials were discarded to...Salud, the Laboratorio de Salud Publica, and the Comité de Investigaciones in Peru gave written consent for the trials and associated protocols. Sra

Control of aedine mosquito vectors, either by mosquito population reduction or replacement with refractory mosquitoes, may play an essential role in the fight against arboviral diseases. In this review, we will focus on the development and application of biological approaches, both natural or engineered, to limit mosquito vector competence for arboviruses. The study of mosquito antiviral immunity has led to the identification of a number of host response mechanisms and proteins that are required to control arbovirus replication in mosquitoes, though more factors influencing vector competence are likely to be discovered. We will discuss key aspects of these pathways as targets either for selection of naturally resistant mosquito populations or for mosquito genetic manipulation. Moreover, we will consider the use of endosymbiotic bacteria such as Wolbachia, which in some cases have proven to be remarkably efficient in disrupting arbovirus transmission by mosquitoes, but also the use of naturally occurring insect-specific viruses that may interfere with arboviruses in mosquito vectors. Finally, we will discuss the use of paratransgenesis as well as entomopathogenic fungi, which are also proposed strategies to control vector competence.

Control of aedine mosquito vectors, either by mosquito population reduction or replacement with refractory mosquitoes, may play an essential role in the fight against arboviral diseases. In this review, we will focus on the development and application of biological approaches, both natural or engineered, to limit mosquito vector competence for arboviruses. The study of mosquito antiviral immunity has led to the identification of a number of host response mechanisms and proteins that are required to control arbovirus replication in mosquitoes, though more factors influencing vector competence are likely to be discovered. We will discuss key aspects of these pathways as targets either for selection of naturally resistant mosquito populations or for mosquito genetic manipulation. Moreover, we will consider the use of endosymbiotic bacteria such as Wolbachia, which in some cases have proven to be remarkably efficient in disrupting arbovirus transmission by mosquitoes, but also the use of naturally occurring insect-specific viruses that may interfere with arboviruses in mosquito vectors. Finally, we will discuss the use of paratransgenesis as well as entomopathogenic fungi, which are also proposed strategies to control vector competence. PMID:26463078

Full Text Available Control of aedine mosquito vectors, either by mosquito population reduction or replacement with refractory mosquitoes, may play an essential role in the fight against arboviral diseases. In this review, we will focus on the development and application of biological approaches, both natural or engineered, to limit mosquito vector competence for arboviruses. The study of mosquito antiviral immunity has led to the identification of a number of host response mechanisms and proteins that are required to control arbovirus replication in mosquitoes, though more factors influencing vector competence are likely to be discovered. We will discuss key aspects of these pathways as targets either for selection of naturally resistant mosquito populations or for mosquito genetic manipulation. Moreover, we will consider the use of endosymbiotic bacteria such as Wolbachia, which in some cases have proven to be remarkably efficient in disrupting arbovirus transmission by mosquitoes, but also the use of naturally occurring insect-specific viruses that may interfere with arboviruses in mosquito vectors. Finally, we will discuss the use of paratransgenesis as well as entomopathogenic fungi, which are also proposed strategies to control vector competence.

Abstract Zika virus, already widely distributed in Africa and Asia, was recently reported in two Northeastern Brazilian: State of Bahia and State of Rio Grande do Norte, and one Southeastern: State of São Paulo. This finding adds a potentially noxious virus to a list of several other viruses that are widely transmitted by Aedes (Stegomyia) aegypti and Aedes (Stegomyia) albopictus in Brazil. The pathology and epidemiology, including the distribution and vectors associated with Zika virus, are ...

The number and productivity of larval habitats ultimately determine the density of adult mosquitoes. The biological and physicochemical conditions at the larval habitat affect larval development hence affecting the adult body size. The influence of biological and physicochemical characteristics on the body size of Anopheles gambiae was assessed in Jaribuni village, Kilifi district along the Kenyan Coast. Ten cages measuring 1 x 1 x 1 m (1 m3) with a netting material were placed in 10 different aquatic habitats, which were positive for anopheline mosquito larvae. Emergent mosquitoes were collected daily by aspiration and the wing lengths were determined by microscopy. In the habitats, physicochemical parameters were assessed: pH, surface debris, algae and emergent plants, turbidity, substrate, nitrate, ammonia, phosphate and chlorophyll a content. A total of 685 anopheline and culicine mosquitoes were collected from the emergent cages. Only female mosquitoes were considered in this study. Among the Anopheles spp, 202 were An. gambiae s.s., eight An. arabiensis, two An. funestus, whereas the Culex spp was composed of 214 Cx. quinquefasciatus, 10 Cx. tigripes, eight Cx. annulioris and one Cx. cumminsii. The mean wing length of the female An. gambiae s.s. mosquitoes was 3.02 mm (n=157), while that of An. arabiensis was 3.09 mm (n=9). There were no associations between the wing lengths and the environmental and chemical parameters, except for a positive correlation between wing length of An. gambiae and chlorophyll a content (r = 0.622). The day on which the mosquitoes emerged was not significant for the anopheline (p = 0.324) or culicine mosquitoes (p = 0.374), because the mosquito emerged from the cages on a daily basis. In conclusion, there was variability in production of emergent mosquitoes from different habitats, which means that there should be targeted control on these habitats based on productivity.

Our ultimate objective is to design cost-effective control strategies for Aedes albopictus, the Asian tiger mosquito, an important urban nuisance and disease vector that expanded worldwide during the last 40 years. ?We conducted mosquitolarval surveys from May through October 2009 in the City of Trenton, New Jersey, USA, while performing intensive monthly source-reduction campaigns that involved removing, emptying, or treating all accessible containers with larvicides and pupicides. We exami...

In 2003, a survey at waste management grounds and tire dealerships was conducted to determine the species composition of mosquitoes in tires in southern Manitoba, Canada. Over 25% of the 1,142 tires sampled contained a total of 32,474 mosquito larvae and pupae. Culex restuans made up at least 95% of the larvae collected for each month of the summer. Culiseta inornata and Culex tarsalis reached their greatest numbers in July and August, respectively, though they were never abundant. Ochlerotatus triseriatus was also found but never reached more than 1% of the total larvae collected in any given month. Mosquito prevalence was more than three times greater in August (36.1%) than in June (11.7%). Orientation affected prevalence of mosquitoes in tires: 31.4% of vertical tires (tires standing on their treads) contained mosquitoes, whereas mosquitoes were found in only 18.9% of horizontal tires (tires parallel to the ground). Tires in the eastern region of Manitoba contained mosquitoes more often (61.7%), irrespective of date, than Winnipeg (25.9%), the central region (29.1%), or the western region (19.8%). Mosquito prevalence was similar across three size categories of tires, car tires (18.8%), truck tires (19.8%), and semi-trailer tires (26.7%), though tractor tires (47.8%) contained significantly more mosquitoes than tires in the other categories.

The mosquitoAedes aegypti is the primary vector of dengue virus (DENV) infection in most of the subtropical and tropical countries. Besides DENV, yellow fever virus (YFV) is also transmitted by A. aegypti. Susceptibility of A. aegypti to West Nile virus (WNV) has also been confirmed. Although studies have indicated correlation of codon bias between flaviviridae and their animal/insect hosts, it is not clear if codon sequences have any relation to susceptibility of A. aegypti to DENV, YFV and WNV. In the current study, usages of codon context sequences (codon pairs for neighboring amino acids) of the vector (A. aegypti) genome as well as the flaviviral genomes are investigated. We used bioinformatics methods to quantify codon context bias in a genome-wide manner of A. aegypti as well as DENV, WNV and YFV sequences. Mutual information statistics was applied to perform bicluster analysis of codon context bias between vector and flaviviral sequences. Functional relevance of the bicluster pattern was inferred from published microarray data. Our study shows that codon context bias of DENV, WNV and YFV sequences varies in a bicluster manner with that of specific sets of genes of A. aegypti. Many of these mosquito genes are known to be differentially expressed in response to flaviviral infection suggesting that codon context sequences of A. aegypti and the flaviviruses may play a role in the susceptible interaction between flaviviruses and this mosquito. The bias in usages of codon context sequences likely has a functional association with susceptibility of A. aegypti to flaviviral infection. The results from this study will allow us to conduct hypothesis-driven tests to examine the role of codon context bias in evolution of vector-virus interactions at the molecular level.